Scientific Program

Conference Series Ltd invites all the participants across the globe to attend 3rd International Conference & Exhibition on Tissue Preservation and Biobanking (10 Plenary Forums - 1 Event)
San Francisco, California, USA.

Day 1 :

Keynote Forum

M Zavos

The Andrology Institute of America, USA

Keynote: The evolution and current status of sperm cryopreservation

Time : 9:45-10:10

Conference Series Biobanking 2017 International Conference Keynote Speaker M Zavos photo
Biography:

M Zavos received his BS in Biology-Chemistry in 1970, his MS in Biology-Physiology in 1972 and Education Specialist in Science (EdS) in 1976 from Emporia State University in Emporia, Kansas. He earned his PhD in Reproductive Physiology, Biochemistry and Statistics in 1978 from the University of Minnesota in the Twin Cities, Minnesota. He received the distinguished Alumnus Award and the Graduate Teaching Award from Emporia State University and the Student Leadership Award from the University of Minnesota. He has numerous scientific collaborations nationally and internationally and his publications have appeared in eight languages. He is a Member of the American Society for Reproductive Medicine (ASRM), the American Society of Andrology (ASA), and the European Society for Human Reproduction and Embryology (ESHRE), the Middle East Fertility Society (MEFS), the Japanese Fertility Society, the International Society of Cryobiology Sigma XI, Gamma Sigma Delta and a number of other Scientific and Professional Societies. He has served on a large number of committees for the International Society of Cryobiology, ASRM, MEFS, ESHRE and others.

Abstract:

Semen or sperm cryopreservation (commonly called sperm banking) is a procedure to preserve sperm cells. Semen can be used successfully indefinitely after cryopreservation which normally is kept at a very stable temperature of -196ºC (liquid Nitrogen). For human sperm, the longest reported successful storage which yielded a successful pregnancy is 22 years. The cryopreservation procedure can be used for sperm donation where the recipient wants the treatment in a different time or place, or as a means of preserving fertility for men undergoing vasectomy or other treatments that may compromise their fertility, such as chemotherapy, radiation therapy or surgery. Cryopreservation extends the availability of sperm for fertilization; however, the fertilizing potential of the frozen-thawed sperm is compromised because of alterations in the structure and physiology of the sperm cell. These alterations, characteristics of sperm capacitation, are present in the motile population and decrease sperm life-span, ability to interact with the female tract, and subsequent fertilizing ability. The etiology of such alterations may represent a combination of factors, such as inherited sensitivity of the sperm cell to withstand the cryopreservation process and the semen dilution. Although the former is difficult to address, approaches that make-up for the dilution of seminal fluid may be sought. The aim of this work is to review aspects of sperm cryopreservation paralleled by events of capacitation and evaluate the possible roles of sperm membrane cholesterol, reactive oxygen species, and seminal plasma as mediators of cryopreservation effects on sperm function. As far as the methods of cryopreservation of human sperm, there are three main conventional freezing techniques used in sperm cryopreservation: Slow freezing, rapid freezing and vitrification. The slow freezing technique which consists of progressive sperm cooling over a period of 2–4 hours in two or three steps, either manually or automatically using a semi programmable freezer; The rapid freezing technique which requires direct contact between the straws and the nitrogen vapors for 8–10 min and immersion in liquid nitrogen at −196°C and lastly, the vitrification method which is the newest procedure to be followed and which renders very high cooling and warming rates (over 20,000°C/min) and short contact with concentrated cryoprotective additives (less than 30 sec over −180°C) and which also offers a possibility to circumvent chilling injury and to decrease toxic and osmotic damage. All three of those methods are clinically employed and some of their technical aspects will be evaluated and compared along with their implications on the outcome of clinical results.

Keynote Forum

Heiko Zimmermann

Fraunhofer Institute for Biomedical Engineering, Germany

Keynote: Improved methods and procedures for pluripotent stem cell preservation, storage stability and validation

Time : 10:10-10:35

Conference Series Biobanking 2017 International Conference Keynote Speaker Heiko Zimmermann photo
Biography:

Heiko Zimmermann is the Director and Head of Fraunhofer IBMT and Chair in Molecular and Cellular Biotechnology at the Saarland University and has been working as Physicist since 1997 in the field of cell biophysics. He coordinated the EU-project HYPERLAB in FP7 and was WP leader in several other EU-projects. Within EBiSC, he is Leader of WP 3 and is additionally coordinating the translational project DropTech® in FP7. He received the first permission for working with hESCs within the Fraunhofer Gesellschaft. He is the author of more than 70 peer-reviewed papers and book chapters. His research expertise covers cryobiology, cryotechnology and biopolymers for clinical scaffolds. He is inventor of more than 50 granted patent families from which more than 20 have been commercially licensed.

Abstract:

The project EBiSC aims to build up a European biobank for research grade human induced pluripotent stem cells (hIPSCs). The vision of EBiSC leads to the demand for upscaled production methods, these kinds of cells leading to the need for automated systems and procedures in stem cell processing and banking. An overview of existing state-of-the-art automation systems is given and the specifications for different applications are compared. Furthermore, modules and concepts for automated cell identification, pluripotency testing, and viability and functionality tests are drawn and results are shown. Scalable label-free analysis of pluripotent stem cells using quantitative life cell imaging and on-line image analysis is shown. A specialized system, the automated hanging drop technique (DropTech®) is shown. The DropTech system allows fully automated cultivation of hiPSCs on micro carrier using the hanging drop technology and enables applications like the automated Embryonic Stem cell Test (EST) for standardized embryo toxicity tests. The last part of the talk deals with the technology of cryopreservation, banking and validation frozen samples. The method of surface-based vitrification of pluripotent stem cells is introduced and the need for a completely closed cool chain is derived from experimental results. Solutions for automated industrial scale biobanking with closed cool chains and with minimal harmful thermal fluctuations are shown and the effect on functionality of cryopreserved cells compared to standard technology is shown. A method for non-invasive monitoring of re-crystallization and de-vitrification effects using Raman micro-spectroscopy is presented.

Keynote Forum

Igor Katkov

Belgorod National Research University, Russia

Keynote: Cryopreservation by vitrification: Basic thermodynamic principals, methods and devices

Time : 10:35-11:00

Conference Series Biobanking 2017 International Conference Keynote Speaker Igor Katkov photo
Biography:

Igor I Katkov is a trained Biophysicist with more than 30 years of experience in Cryobiology and Cryogenic Engineering. His research has been focused on the fundamental aspects of kinetic vitrification (K-VF) as well as on designing the practical system for K-VF KrioBlast™ in cooperation with V F Bolyukh from Ukraine. He is a Chief Scientific Officer of Celltronix, San Diego, USA and recently accepted a Professor-level position of the Head of the Laboratory of Amorphous State in the Belgorod National Research University, Russia.

Abstract:

Cryopreservation (CP) and subsequent long-term storage (cryobanking) are important parts of both life science research and related industries and technologies. As we have stated before, there are 5 basics ways of achieving long-term storage, all essentially lead to vitrification of cells, namely: Slow freezing (SF), Equilibrium vitrification (E-VF) with high concentration of vitrificants (“thickeners”) and relatively moderate speed of cooling and rewarming, Kinetic vitrification (K-VF) with very rapid rates of cooling and rewarming and low to none concentration of exogenous vitrificants, Freeze-drying (lyophilization), and Vacuum/air flow drying at temperatures above zero degree Celsius (xeropreservation), which up to now, is the mainstream of the majority of CP technologies. It however requires multi-step protocols, expensive programmable freezers and must be tuned to the particular types of cells, tissues and organs. In this presentation, we will focus on the kinetic vs. equilibrium vitrification. We will compare the mechanisms, analyze phase diagrams, emphasize the role of the Leidenfrost effect (LFE) and ways of reducing up to full eliminating LFE, discuss pros and cons of each methods and present information on basic equipment and accessories for E-KF and K-VF used in different field of biology, reproductive, regenerative and personalized medicine, drug screening, agriculture, conservation of endangered species, medicine and other related disciplines of sciences and industries. And, finally, we will introduce our (solution to hyper-fast cooling and present a short video clip of the novel hyper-fast scalable cooling devices KrioBlast and VitriPlunger and briefly discuss the promising results on K-VF of human sperm, embryonic stem cells and insulin-producing cells using the KrioBlas™ and VitriPlunger systems.

 

Keynote Forum

Charles F Mahl

GenLife Institute for Regenerative Medicine and Stem Cells, USA

Keynote: Prolotherapy the first choice in Regenerative Medicine

Time : 11.20-11:45

Conference Series Biobanking 2017 International Conference Keynote Speaker Charles F Mahl photo
Biography:

Charles F Mahl is a graduate of Case Western Reserve University in Cleveland, Ohio. He has received his MD degree from the Rosalind Franklin School of Medicine in Chicago, Illinois. He did his Residency and Chief Residency in New York at Interfaith Hospital and Vitreo Retinal Fellowship at the University of Oregon. He has completed a Fellowship program in Anti-Aging and Regenerative Medicine by the American Academy of Anti-Aging Medicine, the Living Younger Preventive Aging Program and is certified in Age Management Medicine by The Cenegenics Education and Research Foundation. He is Regenestem trained in both PRP and adipose/bone marrow derived stem cells. He is Board Certified in Integrative Medicine (BCIM) and is a Certified Sports Trainer (CST) and Certified Sports Nutritionist (CSN). He is also a Member of the Advisory Board of Global Stem Cells and Regenestem.

Abstract:

Prolotherapy has been around for almost 80 years, yet is not well known in most musculoskeletal circles. Most orthopedic surgeons and neurosurgeons either have not ever heard of prolotherapy or think it does not work based on never having studied it in residency training programs, their personal bias or their general lack of knowledge about it. The author will explain his discovery of prolotherapy after some 30 years of suffering from pain and his experience and will introduce this fine medical art to the participants. Prolotherapy was the first procedure invented in Regenerative Medicine, followed by PRP-Platelet Rich Plasma injections and then Stem Cell Injections from the bone marrow and/or adipose tissue. Prolotherapy is a simple technique, much less invasive and less harmful as compared to steroid injections and high risk musculoskeletal surgery and often times, when performed by a skilled and experienced prolotherapist can alleviate the chronic pain one is suffering from. The author will explain the physiology of ligaments and tendons in response to prolotherapy and differentiate between 1st line, 2nd line and 3rd line regenerative medicine therapies. Prolotherapy is the first line of defense when it comes to regenerative medicine and should be recommended to patients prior to cortisone injections or invasive surgery. Elite athletes are already aware of its potential and utilize it when injured and or reinjured to heal their painful joints. It is time for prolotherapy to become main stream and no longer be considered alternative medicine. Author will describe the indications for prolotherapy, the techniques utilized and indications for this therapy. Prolotherapy is an extremely successful therapy in experienced hands and offers powerful pain relief and healing, especially to those who cannot afford PRP or Stem Cells and is therefore, a therapy that be part of all physicians training in third world countries and the like where large percentages of the population cannot afford or have no access to PRP or stem cells. For an indigent population, prolotherapy offers a very good treatment for healing and relief of musculoskeletal pain. Any physician doing stem cell therapy (musculoskeletal) should be well trained in prolotherapy prior to using the advanced therapies of performing platelet rich plasma injections (PRP) and bone marrow aspirates for stem cells and liposuctions for adipose derived stem cells. These procedures are very expensive as compared to prolotherapy. You need to have a major knowledge of injection techniques and know your anatomy-bones, ligaments, tendons and muscles in order to put the cells where they need to be. You need to be able to do these procedures via head, heart and hand (palpation) and via ultrasonic guidance and C-arm Fluoroscopy for more precision. Before rushing into doing stem cell therapies, prolotherapy should be tried because it is a much simpler and less invasive technique. Prolotherapy is currently being performed as a precursor to having PRP and stem cell therapy by competent regenerative medicine doctors. A discussion of what is prolotherapy and its uses and what it does is germane to this talk. It is important for physicians to know that often a simple treatment can be performed to accomplish the same result as long as one is experienced and knowledgeable as a prolotherapist.

 

  • Biobanking | Cryopreservation| Vitrification | Biorepository & Biospecimen| Stem Cell | Regenerative medicine| Tissue engineering | Rejuvenation
Location: Plaza I

Chair

Panayiotis Zavos

The Andrology Institute of America, USA

Co-Chair

Debra Aub Webster

Cardinal Health Regulatory Sciences, USA

Speaker
Biography:

Lourdes Cortes-Dericks completed her PhD in Biological Sciences from the University of Hamburg, Germany. Her dissertation was focused on the cellular and molecular mechanisms of the desensitization processes in guanylyl cyclase receptors. Her postdoctoral studies principally surround the tumor biology of lung cancers and identification of tumor biomarkers for tumor cancer stem cells to identify their roles in lung carcinogenesis. She is an active peer reviewer in numerous respected journals and an editorial board member.

Abstract:

Mesenchymal stem cells (MSCs) are adult stem cells having the capacity for self-renewal and differentiation. The current consensus of the therapeutic benefits of mesenchymal stem cells posits that these cells secret trophic factors that elicit paracrine actions for cancer therapy. These soluble factors may be collected in what has been known as the conditioned medium (CM). In the malignant setting, the biomolecules secreted by MSCs are thought to either support or inhibit tumor growth. This presentation will cover the anti-tumor effect of the human lung mesenchymal stem cell-derived conditioned medium (hlMSC-CM) in lung cancers. It will include the identification of the different biomolecules contained in the hlMSC-CM and its anti-tumor effects in malignant mesothelioma cell lines. It will highlight the efficacy of hlMSC-CM as compared to twice the IC50 of cisplatin in eliminating the chemo resistant, sphere-forming mesothelioma cells. Other regenerative capacities MSC-CM from our and other studies such as the attenuation of cigarette smoke-induced injury in lung fibroblasts, in vivo and in vitro will also be mentioned to underscore its potential in improving tissue regeneration.

In the clinical setting, the use of cell-free MSC-derived conditioned medium may facilitate a practical approach compared with stem cell-based therapy as the former is easier to prepare, preserve and transport to appropriate clinics, and may also present less complications relative to issues on cell transplantation. Despite the promising results of the beneficial effects of MSC-derived conditioned media, standardization of MSC cultivation, collection and preservation of the CM is still necessary. 

Xu Han

CryoCrate LLC and University of Missouri, USA

Title: Life in Nano Ice : Application of cryocrate C80EZ medium for cell and tissue cryopreservation

Time : 12:30-12:50

Speaker
Biography:

Xu Han has over 10 years of experience in the field for Cryopreservation. He is the Top Reviewer of the journal of Cryobiology, and a winner of NIH SBIR, Coulter Translational Partnership Program Award, and MU Faculty Innovation Award. He has endeavored in improving cryopreservation methods and devices. His expertise is also in the fields of Calorimetry, Numerical Simulation, Thermodynamics and Mechanics.

Abstract:

Traditional cryopreservation approaches are technically problematic in either producing large ice crystals (typically on um scale) associated with equilibrium freezing process or involving extremely high concentrations (40-60% v/v) of permeating cryoprotectants to achieve vitrification. Both also require the use of liquid nitrogen and expensive facilities for long-term storage or banking. With the support of NIH SBIR, Coulter Translational Partnership Program, and University of Missouri (MU) Fast-track Award, CryoCrate (a MU spin-off) has developed an innovative cryopreservation medium, trademarked as C80EZ, and opened the door for cryopreservation with both nano scale ice formation and long-term storage in regular deep freezers at approximately -80ºC. The base medium of C80EZ enables long-term storage of cells at -80ºC by preventing recrystallization and the efficiency has been demonstrated by previous publications, e.g. Sci Report 2016 (nature.com). The complete C80EZ medium enables unique nano scale ice formation and the efficiency tests have been performed on cell or tissue types that survive poorly from any traditional cryopreservation method, e.g. Human corneas, primary neurons, porcine and rodent spermatozoa, and cell types that cannot be stored in deep freezers for extended periods of time, e.g. prophetical blood mononuclear cells and mouse embryos. Significant improvement of cell post-thaw viabilities and functionalities has been observed for all above cell and tissues types. For E. coli competent cells (routinely stored in deep freezers), our preliminary data showed that the nano ice technology can remarkably increase post-thaw transformation efficiency. Currently, the C80EZ medium doesn’t improve survivability from cryopreservation for cell types with large volume of intracellular lipid components, e.g. porcine and fish embryos. The C80EZ medium is now commercially available, and the technology is patent pending (PCT filed) and exclusively licensed from MU to CryoCrate.

Speaker
Biography:

Dr. Jedediah Lewis currently serves as Chief executive officer at Organ Preservation Alliance.  Dr. Jedediah Lewis holds degree from Harvard law school and also work as a research assistant in Harvard law school and Emory university school of medicine.

Abstract:

A growing research effort aims to achieve the cryopreservation of viable large tissues and organs for both transplantation and research, with the potential to save or improve the lives of millions of patients worldwide. Recent years have seen multiple targeted federal grant pipelines for complex tissue cryobanking that have funded dozens of labs across the U.S., as well as roundtables at the White House and on Capitol Hill, an NSF-funded technology road mapping process, and other meetings with U.S. science agencies and stakeholder organizations to generate support for organ and tissue banking progress. At two global Organ Banking Summits, held at sites including Stanford School of Medicine and Harvard Medical School, leading cryopreservation labs have mapped out the remaining “sub-challenges” to cryo bank whole organs and large tissues in a viable state. The American Society of Transplantation recently launched a community of practice that aims to support advances toward the cryopreservation of whole organs, limbs, and an array of other biological systems. These developments offer numerous opportunities for the biobanking community, as they can enable both new applications of bio banked tissue, new cold chain solutions for cells and tissues, and new support for biobanking advances.

Speaker
Biography:

Yulian Zhao is an Associate Professor of Gynecology and Obstetrics and Director of Assisted Reproductive Technology Laboratory at the Department of Gyn/Ob, the Johns Hopkins University School of Medicine. She is an Officer, Board Member, Committee Member and Active Member in a number of professional organizations and societies in the field of Reproductive Medicine. She frequently acts as an ad hoc reviewer for over 12 professional journals and has been a recipient of several academic awards. She has authored over 50 peer-reviewed articles, review articles and book chapters as well as delivered over 50 presentations at the professional conferences and society events nationally and internationally.

Abstract:

Loss of fertility can significantly compromise quality of life. For populations of reproductive age, fertility preservation strategies have been well established and widely used, but most of the technologies are not options for pediatric age group, because of their sexual immaturity. Majority of such cases would be cancer patients. One of the medical late effects of cancer treatment is infertility or severely compromised fertility. Fertility preservation is of great importance to psychological well-being and to the quality of life for these patients. Fertility preservation for this population has become an issue of attention in recent decades. Strategies for long-term preservation and storage of reproductive tissue for this particular population are of key importance for scientific research and reproductive medicine. This emerging field encompasses cryopreservation of reproductive cells including testicular cell suspensions and testicular tissue. There are 3 potential strategies under active investigation. The first one is to cryopreserve testicular tissue or cell suspension before cancer therapy. Once the patient is cured from cancer and ready to begin a family, the tissue or cells could be thawed and re-implanted into the patient's own testes to continue full maturation. The second option is in vitro differentiation of testicular tissue. Spermatozoa may be generated from spermatogonial stem cells in vitro until they can be used for in vitro fertilization or be re-implanted into the testes. The third one is that immature cryopreserved testicular tissue may be grafted into another organ or tissue of the patient. These fertility restoration techniques have been successfully applied in several animal models and are considered to be very promising for future clinical applications in human.

Jill Davies

Oxford University, UK

Title: Fertility cryopreservation in Oxford, UK

Time : 14:30-14:50

Speaker
Biography:

Jill Davies graduated from Coventry University in 1987 with a degree in Applied Biology. After university studies, she worked for cardiothoracic surgeons Mr. Donald Ross & Sir Magdi Yacoub at the National Heart Hospital in London and that she opened the heart valve bank at the Oxford University Hospital in Oxford, England in 1990. The Oxford bank supplies cardiovascular tissue, corneas for transplant and research and brains & spinal cords for research. It is now also an Oxford Research Center Bio bank. She is also an executive member of the BATB, member of AATB, SLTB, and Society for Cryobiology.

 

Abstract:

Late effects of cancer treatment may cause premature infertility in children and adolescent cancer survivors. Oxford applied for UK license for cryopreservation of ovarian and immature testicular tissue (ITT) for patients requiring gonadotoxic treatment. This required validation with animal/human studies, development of protocols, information, staff training, establishing costs/funding source. Oxford recruited first set of patients in 2013. Patients eligible for treatment intent is curative, cytoxicity high risk, age <40 years (females) and boys pre-pubertal. Patients consented prior to day of surgery. Ovarian tissue from 145 females, age 10 months – 39 years cryopreserved (14% with benign diagnosis), testicular tissue (ITT) from 31 boys, (0.9–13.6 years). Referrals from oncologists/hematologists and patients, most patients travel to Oxford for surgery scheduled to coincide with other interventions (minimizing risk of anesthesia). Right ovary procured using laparoscopic oophorectomy, wedge testicular biopsy procured via midline incision. Tissue bank technician collects tissue and dissects within cleanroom, producing ovarian cortical strips (1x1x5mm) containing primordial follicles or testicular segments (2x2x2 mm) containing spermatagonial stem cells. Histological evaluation reviews viability and presence of metastatic cells. Immature oocytes collected for in vitro maturation (IVM) and vitrified. Cryopreservation of ovarian tissue requires ethylene glycol, sucrose, human albumin serum, slow controlled freezing (manual seeding at 8ËšC) and vapor phase nitrogen storage. DMSO is used for ITT. Tissue is stored until the patient is cured and confirmed infertile. 86 live births following ovarian tissue transplantation was recorded. A live birth in ITT is successful in animal studies only. Patients with hematological malignancy will use vitrified oocytes (gametes are free from residual disease). For other females and ITT patients, tissue will be matured in vitro (IVM) to develop primordial follicles/spermatogenesis for use in conventional ART. Oxford University researchers currently are optimizing IVM techniques (three dimensional/organ culture studies). All details were registered in OTCP database. Service evaluation (interviews with patients/parents) confirms patients/parents understood risks/potential for fertility restoration. Setting up the Oxford service has been challenging due to evolving UK legislation. Referrals are from all UK centers.

 

Mohamed A Zayed

Washington University School of Medicine, USA

Title: Peripheral artery and blood biobanking can lead to scientific collaboration and discovery

Time : 14:50-15:10

Speaker
Biography:

Mohamed A Zayed is Surgeon-Scientist at the Department of Surgery, Section of Vascular Surgery, and Washington University School of Medicine, USA. He has completed his Medical training at Stanford University and earned his Doctoral degree in Pharmacology at the University of North Carolina at Chapel Hill. He has previously served as the Chief Medical Officer for a software start-up company and has published over 30 research articles in reputable journals. His current clinical and research interests focus on the influences of diabetes on peripheral arterial disease.

Abstract:

Objectives: Over 3 million Americans have advanced peripheral arterial occlusive disease leading to significant patient morbidity and mortality. The lack of well-preserved human peripheral arterial tissue substrate has limited scientific exploration of this disease process and development of impactful-targeted molecular therapies. To address this, we developed an integrative biobanking strategy to collect peripheral arterial tissue specimens from patients undergoing vascular surgery.

Methods: Over 29 months, we harvested vascular specimens from consenting patients undergoing open arterial endarterectomy and revascularization procedures. All patients were enrolled in an IRB approved protocol. A biobank infrastructure was developed to manage logistics, funding, collection, and real-time processing of harvested arterial tissue.

Results: 486 patients were enrolled in the vascular surgery biobank prior to their index operation. Forty-two (42) clinical variables were evaluated for each patient during the perioperative period. Vascular specimens were successfully collected for 72.3% (349) of patients who enrolled in the biobank. The majority of specimens collected were retrieved from the peripheral arterial system (38.7% carotid artery, 9.5% anterior or posterior tibial arteries, 24.4% femoral or popliteal arteries). For all patients, blood samples were collected and processed to provide serum (85.5%) and plasma (86.1%). Each arterial specimen was sub-divided into maximally and minimally diseased portions to facilitate intra- and inter-patient biochemical and molecular analyses. Over the study period 8 collaborations (in 4 different departments at 2 universities) were fostered to provide 82 tissue specimens and 98 blood samples.

Conclusions: An integrative biobanking approach in vascular surgery patients is feasible and provides a highly unique peripheral arterial substrate for molecular and biochemical analyses. Biobanking management and daily operations requires a dedicated team approach to insure proper patient consenting, specimen collection, subsequent experimental analysis and meaningful scientific collaborations.

Dalip Sethi

Cesca Therapeutics Inc., USA

Title: Clinical applications of autologous bone marrow derived cells

Time : 15:10-15:30

Speaker
Biography:

Dalip Sethi currently serves as Director of Clinical Research at Cesca Therapeutics Inc., a public company engaged in the research, development, and commercialization of cellular therapies and delivery systems for use in regenerative medicine. He holds a PhD in Biotechnology from Institute of Genomics & Integrative Biology, New Delhi, and a Master’s in Organic Chemistry from Hansraj College, Delhi University, India, and conducted Post-doctoral studies at Thomas Jefferson University, School of Medicine. He is currently engaged in the development of point-of-care devices, methods and diagnostics for use in autologous cell therapy in the treatment of cardiovascular and orthopedic diseases.

Abstract:

Cardiovascular diseases are a major burden on healthcare in modern society. Diseases, such as Critical Limb Ischemia (CLI), are debilitating. Many of the cardiovascular ischemic disease patients have limited surgical or medical options. Regeneration of vascular system is an attractive treatment strategy and is actively pursued in various preclinical and clinical settings. One of the options in regenerative medicine is the use of autologous bone-marrow concentrate (aBMC) containing stem and progenitor cells. Autologous bone-marrow concentrate (aBMC) is derived from the bone marrow aspirate (BMA) by density centrifugation and can be delivered either intra-muscularly (IM) or intra-coronary in the affected region. The aBMC consists of a) an acellular fraction comprised of autologous plasma and the cytokines and b) cellular fraction which is a source of (i) proangiogenic cells such as hematopoietic stem cells, mesenchymal progenitor cells, and endothelial progenitor cells; (ii) other cells of immune system at different levels of maturity and multi-potency. The acellular and cellular components participate in tissue repair and regeneration and have made aBMC an attractive source of cells and cytokines for therapeutic angiogenesis in the treatment of ischemic diseases.

Speaker
Biography:

Raffaella Fabbri has been working as a Biologist in the Human Reproductive Medicine Unit, S Orsola-Malpighi Hospital of University of Bologna in Italy since 1977. In 2001, she received a national and international worldwide patent about method and solution for cryopreserving oocytes. In 2006, she obtained the United States Patent US 7,011,937 B2 for method and solutions for cryopreserving oocytes. She has her expertise in cryopreservation of human mature and immature oocytes, cryopreservation of embryos and blastocysts, cryopreservation of human ovarian tissue in vitro maturation of ovarian follicles, co-cultures of human granulosa cells and sperms, fertilization and embryonal development. She has been involved in many scientific activities which are well-documented by 335 articles published in Italian and foreign journals, participated in 134 meeting as speaker and 58 as Auditor.

 

Abstract:

Statement of the Problem: Ovarian tissue cryopreservation represents a valid strategy to preserve ovarian function in cancer patients with a high risk of premature ovarian failure due to chemo/radiotherapy. The ovarian tissue remains frozen for very long period of time (the request of tissue replanting usually occurs after at least five years from the end of therapies and this period may dragging on further in the case of diseases that require prolonged treatments or in the case of pediatric patients). The purpose of this study is to evaluate the morphology and functional activity of cryopreserved ovarian tissue stored for 18 years after thawing and transplantation.

Methodology & Theoretical Orientation: Ovarian tissue of a 29 year old patient suffering from Hodgkin Lymphoma was cryopreserved at our Centre before starting anticancer treatment. 18 years after storage; her ovarian tissue was evaluated by light microscopy, transmission electron microscopy, TUNEL assay and LIVE/DEAD viability/cytotoxicity test and then heterotopically transplanted in two subcutaneous pockets of patient. Follicle development was evaluated by ultrasound examination on the graft sites.

Findings: Ovarian tissue showed a good morphology, no apoptosis signs, sub-cellular integrity of follicles and interstitial edema foci. The LIVE/DEAD assay performed on stromal cells, isolated from cryopreserved tissue, showed viable cells (>97%) after 2 and 7 days of culture. The patient had the first menstruation five months after transplantation and to date (20 months from the graft), she is regularly menstruating every 30-40 days. Follicular development is monthly evidenced by a bulge palpable beneath the skin in the graft sites.

Conclusion & Significance: This study provides evidence that the storage time does not impact on tissue quality and on tissue ability to resume the ovarian function after replanting. These results give hope especially to cancer girls, whose tissues could remain cryopreserved for a very long time.

 

Achour Radhouane

El-Manar University, Tunisia

Title: Premature ovarian failure diagnosis and management

Time : 15:30-15:50

Speaker
Biography:

ACHOUR Radhouane is associate professor at faculty of medicine of Tunis-Tunisia; He has published many basic and clinical articles in relation to gynecology and obstetrics. , his research interests include Rare Diseases in gynecology and prenatal diagnosis.

He serves as associate professor, Emergency Department of Gynecology and Obstetrics in maternity and neonatology center Tunis ; Faculty of Medicine of Tunis- El Manar University of Tunis-Tunisia. He also serves as member of the editorial team for: Asian Pacific Journal of Reproduction, the Global Journal of Rare Diseases, Journal of Neonatal Biology, Current pediatric research, Obstetrics and Gynecology: Open access, Pediatrics and Health Research and Member of the Science Advisory Board.

Abstract:

Introduction: Premature ovarian failure (POF) is defined as menopausal levels of follicle stimulating hormone (FSH≥40 IU/l) associated with more than 4 months of secondary amenorrhea occurring before the age of 40. It affects approximately 1% of women and the underlying an etiology remains very complex and heterogeneous. Recently Welt suggested changing the term POF to POI, which stands for premature ovarian insufficiency, because this better reflects the longitudinal progression towards the final menstrual cycle.

Objective: The objective of this study was to investigate different etiologic mechanisms of premature ovarian failure.

Material & Methods: A retrospective study of 30 cases of early menopause reported in our service for period of 10 years from January 2000 to December 2009. We will center this work to draw up an epidemiologic profile of our population, to specify the various circumstances of discovery, to analyze the various mechanisms etiopathologic and finally to discuss the therapeutic possibilities.

Results: The incidence of premature ovarian failure is around 1 to 3%. This pathology occurs in young women who often wish to become pregnant. Two mechanisms could be involved: Initial follicle depletion and follicle dysfunction; however in some cases mixed mechanisms are involved initially. Our study includes 30 patients. Genetic anomalies were found in 3 cases (10%). Anti-ovarian antibodies were found in 18 cases (60%). Ovarian biopsy done in 20 patients confirms immunological mechanism of ovarian failure in 16 cases (70%) with autoimmune ovaritis. In 9 cases idiopathic ovarian failure was observed.

Discussion: The POF etiology in women with normal karyotype remains poorly understood. However, more and more genes involved in the normal follicular and ovarian development are becoming known, which are also likely to be involved in this disease, based on the data obtained from animal models. In such context, the first study presented herein was conducted to evaluate whether the ovarian biopsy, when compared with data provided by pelvic ultrasound (US), could be a reliable tool to individualize patients with an ovarian phenotype suggestive of a particular gene mutation, therefore, suggesting the usefulness of laparoscopy and ovarian histology in orientating the search for the possible genetic etiologies of the POF syndrome. In our experience, when women had no follicle at US, histological examination did not detect follicles in most of our patients. At the contrary, only 56% patients with normal size ovaries with the presence of follicles suggested at US, displayed follicles when histological examination of an ovarian biopsy was performed. Consequently, it can be assumed that, US is not predictive of the presence of follicular structures within the ovary.

Conclusion: The proportion of idiopathic premature ovarian failure tends to decrease owing to the better recognition of genetic and immunological anomalies. Finally the use of the ovarian biopsy still remains interesting in our hands to help us to better understand the process of ovarian dysfunction and is mandatory in patients for whom anomalies of genes involved either in follicular growth or maturation are found.

Brian M Mehling

Blue Horizon International, USA

Title: Mesenchymal stem cells in clinical applications

Time : 16:30-16:50

Speaker
Biography:

Brian M Mehling is a practicing American Orthopedic Trauma Surgeon, Researcher, and Philanthropist. He started his path in Medicine through Undergraduate study at Harvard University, obtaining Bachelor of Arts and Master of Science degrees in Biochemistry from Ohio State University. Completing his degree of Medicine at Wright State University School of Medicine, he received Post-graduate education through residencies and fellowships at St. Joseph’s Hospital in Paterson, NJ and the Graduate Hospital in Philadelphia, PA, while pursuing a PhD in Chemistry. He operates his own practice, Mehling Orthopedics, in both West Islip, NY and Hackensack, NJ.

 

Abstract:

Mesenchymal stem cells (MSCs), multipotent adult stem cells feature the potential to regenerate tissue damage and inhibit inflammation. Our research studies designed to measure the safety and efficacy of intra-venous, intra-articular, and intra-thecal stem cell therapies. Retrospective Chart Review study of umbilical cord blood (UCB) stem cell therapy for spinal cord injury showed that significantly higher proportion of patients in the stem cell therapy group showed improved function in pain and temperature sensation, lower limb muscle strength, bladder and gastrointestinal function compared to the traditional therapy group. UCB stem cell therapy of patients with Sequelae of cerebro-vascular hemorrhage and CVA (Stroke) Sequelae showed significant improvement in mobility and muscle strength of upper and lower extremities, improvement of neurological function. In the period from 2015 to 2016, 207 subjects with musculo-skeletal conditions underwent the therapy with their own stromal vascular fraction cells. Analysis of subjects’ pain sensation and mobility showed the following: 10 days after the therapy, pain sensation decreased by 26.8%. Three months after therapy, the number of subjects with a decrease in pain sensation increased to 50.0%, and six months later, this number was 52.5%. Analysis of subjects’ mobility showed the following: 10 days after the therapy, mobility was improved by 26.0%. Three months after therapy, the number of subjects with improved mobility increased to 43.8%, and six months later this number was 47.4%. Statistical analysis showed that the decrease in subjects’ pain sensation and improvement of subjects’ mobility three and six months after the therapy was statistically significant.

 

Speaker
Biography:

Kay Ohlendieck has an Undergraduate degree in Biology from the University of Konstanz, Germany (1985), a PhD in Biochemistry from University College Cork, Ireland (1989) and a DSc in Muscle Biology from University College Dublin, Ireland (2011). He has worked as a Post-doctoral Associate at the University of Iowa, Iowa City and at the State University of New York, Stony Brook, as well as a Lecturer in the Department of Pharmacology, University College Dublin (1995-2001). Since 2002, he is Chair of Biology at the National University of Ireland, Maynooth, and his research focuses on skeletal muscle proteomics

Abstract:

Statement of the Problem: The X-linked neuromuscular disorder Duchenne muscular dystrophy is caused by primary abnormalities in the dystrophin Dmd gene and is associated with progressive skeletal muscle wasting, sterile inflammation and reactive myofibrosis. The purpose of this study was to analyze tissue degeneration and regeneration in different muscle subtypes that exhibit varying degrees of pathophysiological susceptibility to fiber necrosis.

Methodology & Experimental Approach: A comparative mass spectrometry-based proteomic analysis was carried out to determine distinct alterations in various skeletal muscles from the dystrophic mdx and mdx-4cv animal models. This included the characterization of muscle proteins in mildly versus severely affected skeletal muscles, as well as the label-free mass spectrometric evaluation of proteome-wide changes following fiber regeneration. The independent verification of proteomic findings was carried out by comparative immunoblotting surveys with isoform-specific antibodies to key muscle protein markers and immune-fluorescence microscopy.

Findings: In this study, the complexity of changes in the skeletal muscle proteome during tissue degeneration and regeneration could be established. Muscle-associated proteins belonging to the contractile apparatus, excitation-contraction coupling mechanism, metabolic pathways and the cellular stress response were shown to undergo changes in their abundance and/or isoform expression pattern. Contractile proteins, molecular chaperones, mitochondrial enzymes, cytoskeletal proteins and components of the extracellular matrix exhibited differential changes in moderately versus severely dystrophic fiber populations. During regeneration cycles, the reversal of a variety of protein changes was established by comparative proteomics.

Conclusion & Significance: The newly identified proteomic markers of tissue degeneration and regeneration might represent suitable biomarker candidates to advance our general understanding of the molecular pathogenesis of muscular dystrophy, as well as being useful for improving the diagnosis, prognosis and therapy monitoring of dystrophinopathies

Debra Aub Webster

Cardinal Health Regulatory Sciences, USA

Title: Regenerative medicine–from promise to patient

Time : 17:10-17:30

Speaker
Biography:

Debra Aub Webster has over 20 years of experience in Pharmaceutical Research and the Regulatory Environment. She started her regulatory career with the US FDA as Reviewing Toxicologist/Pharmacologist. As a Principal Scientist in Regulatory Affairs and Product Development with Cardinal Health Regulatory Sciences, she is the Project Lead for biologic and regenerative medicine product development programs. In this capacity, she provides guidance on clinical, nonclinical, and regulatory aspects of strategic product development, author regulatory documents, and acts as the regulatory representative for sponsors in interactions with FDA.

Abstract:

The expansion in the use of human cells and tissues from their functions in practice of medicine, for repair or replacement, to their use as a manufactured product intended to treat an indication or condition has led to increased regulatory oversight by FDA. Under the authority of Section 361 of the PHS Act, FDA introduced a comprehensive regulatory program in 1997 for human cells, tissues, and cellular and tissue-based products (HCT/Ps). This consolidated regulatory approach is tiered and risk-based to allow for less regulatory evaluation of products determined to present a minimal risk to patient safety. In 2005, this regulatory program was implemented in rules codified under 21 CFR Section 1271. Under these rules, some HCT/Ps are defined as biological products, requiring market clearance. The potential regulatory pathways for HCT/P-derived technologies is often complex, and understanding these regulatory pathways and the definitions surrounding them is critical to successful product development. Navigating the regulatory path, from scientific discovery to initiation of clinical investigations is challenging. These challenges include determination of the appropriate regulatory pathway and understanding the requirements regarding demonstration of safety and efficacy and identification of appropriate quality attributes regarding product safety, quality, and potency. FDA has issued numerous guidance documents and offers several expedited approval pathways in order to bring effective discoveries safely forward, to benefit patients. Successful strategies for achieving this goal include identifying critical information gaps with respect to the appropriate regulatory pathway; informed clinical, non-clinical, and quality development plans; maintaining a Target Product Profile; and successful interaction with the appropriate review division within FDA.

 

Speaker
Biography:

Robin C Muise-Helmericks is currently an Associate Professor in the Department of Regenerative Medicine and Cell Biology, Member of the Hollings Cancer Center and Adjunct Associate Professor in the Departments of Oral Health at the Medical University of South Carolina.

Abstract:

The structural balance between elastin, collagen and other extracellular matrix components is absent in adult wound repair resulting in misaligned and excessive collagen deposition that contributes to scar formation. Our findings show that a marine derived pGlcNAc nanofiber stimulates tensile strength and elasticity of wounded skin so that it is equal to that of uninjured skin. Nanofibers promote cellular alignment, secretion of aligned collagen fibers and increased tropoelastin expression. Nanofiber-dependent reductions in scar formation are, in part, due to activation of an integrin/Akt1-dependent pathway. In addition to the activation of Akt1, our deep sequencing results suggest a role for a TLR4/type-I interferon (IFNαβ) pathway in the nanofiber-driven anti-fibrotic phenotype. Here we show that pGlcNAc nanofibers specifically stimulate type-I IFN (IFNαβ) expression via activation of Toll-like receptor 4 (TLR4), in the absence of TLR4-induced NFκB-dependent inflammatory responses in normal human fibroblasts and endothelial cells. TLR4 null animals fail to respond to nanofiber treatment, which can be rescued by addition of IFNα2A to the wound bed. Indeed, inhibition of type-I IFN activity using a blocking antibody against the type-I IFN receptor inhibits nanofiber induced tissue repair. Our findings support a model where nanofiber stimulation of TLR4 preferentially favors the IFNαβ response over an inflammatory response mediated through MyD88. As inflammatory responses lead to increased TGF-β production, increases in myofibroblast production and fibrotic, disorganized collagen deposition. Our findings suggest that reductions in inflammatory cell recruitment or activation results in reduced scar formation and increased tensile strength and elasticity of healed wounds. The marked difference in collagen deposition and increased elastogenesis in nanofiber treated wounds suggests that the nanofibers are specifically stimulating a more regenerative type of tissue repair.

 

Speaker
Biography:

Dr. Osorio brings over 20 years of experience in medical practice.  Served in capacities in private and public practices, as hospital staff physician and as emergency health services coordinator for a variety of private and public institutions throughout Mexico and managed a blood bank from 2009- 2012.  Earned MD degrees at Westhill University and the National Autonomous University of Mexico as a Medico Cirujano. In addition, has diplomas in aesthetic medicine from the Autonomous University of Guadalajara, as an Advance Fellow by the American Board of Anti-Aging and Regenerative Medicine, visiting scholar at University of North Carolina at Chapel Hill in dermatology. Fellow of Stem Cell Medicine by the American Academy of Anti-Aging Medicine and University of South Florida, and currently completing additional masters work in metabolic and nutrition sciences at University of South Florida and Master in Health Sciences Aged Services, Victoria University at Melbourne Australia. A member of the Round table of the ReGeNeRaTe laboratories Mexico’s Committee, a DNAge-Lab Company,  working on the stem cells field since 2007, and since 2011 a member of the International Cellular Medicine Society. Chief Medical Officer & Co-Founder at Astrum Salud Móvil in Mexico City. PRP certified practitioner in the aesthetic and regenerative field since 2011 by GyCo.

Abstract:

We have previously demonstrated that co-electroporation of Xenopus laevis frog oocytes with normal cells and cancerous cell lines induces the expression of pluripotency markers, and in experimental murine model studies that Bioquantine® extract (purified from intra- and extra-oocyte liquid phases of electroporated oocytes) showed potential as a treatment for a wide range of conditions Squint, Spinal Cord Injury (SCI), Cerebral Palsy and skin wrinkling. The current study observed beneficial changes with Bioquantine® administration in a small group of patients with a variety of degenerative disorders. Pluripotent stem cells have therapeutic and regenerative potential in clinical situations including dermatological and CNS disorders even cancer. One method of reprogramming somatic cells into pluripotent stem cells is to expose them to extracts prepared from Xenopus laevis oocytes. We showed previously that co-electroporation of Xenopus laevis frog oocytes; with normal cells and cancerous cells lines, induces expression of markers of pluripotency. We also observed therapeutic effects of treatment with a purified extract (Bioquantine) of intra- and extra-oocyte liquid phases derived from electroporated X. laevis oocytes, on experimentally induced pathologies including murine models of melanoma, traumatic brain injury, and experimental skin wrinkling induced by squalene-monohydroperoxide (Paylian et al, 2016). The positive human findings for Squint, Spinal Cord Injury, Cerebral Palsy and wrinkling with the results from previous animal studies with experimental models of traumatic brain injury, skin wrinkling, and melanoma,respectively (Paylian et al,2016). Because of ethical reasons, legal restrictions, and a limited numbers of volunteers, we were able to treat only a very small number of patients. These results indicate that Bioquantine® may be safe and well tolerated for use in humans, and deserves further study in a range of degenerative disorders. We propose that the mechanism of action of Bioquantine® in these various diseases derives from its unique pharmacology and combinatorial reprogramming properties. In conclusion, these preliminary findings suggest that Bioquantine is safe and well tolerated and contains ubiquitous therapeutic activity including restoration Squint (extropia),Cerebral Palsy, dermatologic healing, spinal cord injury.

Nan Xiao

University of the Pacific, USA

Title: Glial cell derived neurotrophic factor promotes dental pulp stem cell migration

Time : 18:10-18:30

Speaker
Biography:

Nan Xiao is an Assistant Professor of the Department of Biomedical Sciences, Arthur A. Dugoni School of Dentistry, University of Pacific. She is a member of the American Dental Education Association (ADEA) and International Association of Dental Research (IADR). She serves as editor for multiple scientific journal. She also works intensively to promote the dental academic career pathways for dental students. Her academic interest include research in application dental pulp stem cells mediated tissue regeneration and integration the pedagogic innovations in dental education.

Abstract:

Preserving the vitality of the teeth is critical in maintaining the function and esthetics during dental treatment. Dental pulp stem cells (DPSCs) are mesenchymal cells that are demonstrated to possess stem cell properties, such as self-renewal, proliferation and pluripotency. DPSCs can be obtained through non-invasive procedure from the dental pulp and become potential resources for tissue regeneration. Neurotrophic factors are known to promote survival and growth of neurons. In the present study, we examined the expression of the glial cell derived neurotrophic factors (GDNF) family ligands and receptors, and characterized the intracellular localization of them in DPSCs. GDNF family ligands increased the migration of the DPSCs. In addition, we found the AKT and MAPK pathways are downstream of GDNF in regulating the DPSC wound healing and migration. Our results indicate that neurotrophic factors play a role in dental pulp regeneration and may be potential novel therapies for post pulpotomy treatment in adult teeth.

  • Biobanking | Cryopreservation| Vitrification | Biorepository & Biospecimen| Stemcell | Regenerative medicine| Tissue engineering | Rejuvenation
Location: Plaza I

Chair

Heiko Zimmermann

Fraunhofer Institute for Biomedical Engineering, Germany

Co-Chair

Ken yaegaki

Nippon Dental University School of Life Dentistry, Japan

Session Introduction

Aijun Wan

University of California, USA

Title: Engineering stem cells and biomaterials to treat birth defects before birth

Time : 14:20-14:40

Speaker
Biography:

Aijun Wang is the Co-Director of the Surgical Bioengineering Laboratory and an Assistant Professor at the University of California Davis School of Medicine. His research interests center on Engineering Stem Cells and Biomaterials to develop novel regenerative medical therapies, especially surgical treatments for congenital anomalies. His lab has successfully combined Tissue-Engineering technologies with the most advanced fetal intervention, and developed novel biomaterial and stem cell-based treatments (including nanofibrous materials, fetal membrane, decellularized extracellular matrix, iPSC-derived stem cells, placenta-derived stem cells) for devastating birth defects, such as spina bifida. Currently, his team are performing IND enabling studies to move novel stem cell based therapies to clinical applications.

Abstract:

Statement of the Problem: Birth defects are common, affecting approximately 3% of all live births in the United States. Congenital defects can result in significant morbidity and are the most common cause of infant mortality, accounting for a minimum of 20% of all infant deaths worldwide. Improvements in prenatal screening and diagnostics- including ultrasonography, chorionic villus sampling, amniocentesis, fetal magnetic resonance imaging, and, most recently, circulating fetal DNA in maternal blood have allowed earlier diagnosis of a range of congenital disorders. With prenatal diagnosis came the pursuit of prenatal interventions to prevent or address pathology that developed in utero.
 
Methodology & Theoretical Orientation: Our lab has been developing fetal tissue engineering approaches using different types of stem cells and biomaterials to treat a variety of birth defects before birth. One example is in utero transplantation of stem cells with extracellular matrix-based scaffolds to treat spina bifida. Our recent data have shown that treatment with early
gestation placenta-derived mesenchymal stromal cells (PMSCs) during in utero repair cures spina bifida-associated motor function at birth in a fetal lamb model. Based on the previous studies, we are refining the treatments and pursuing IND enabling studies for future clinical translations. We are also developing in utero stem cell transplantation approaches to treat other birth defects such as hemophilia, tracheal occlusion and congenital diaphragmatic hernia.
 
Conclusion & Significance: Stem cell therapies have the potential for use in fetal Tissue Engineering applications, as a vehicle for fetal Genetic Engineering, for the induction of fetal tolerance for postnatal transplantation, and perhaps even for use in fetal treatment of potential adult disease. The fetal environment is ideal for the introduction of cell-based therapies prior to
development of pathology. As with any new therapeutic modality, the ideal technique, potential risks, and long-term efficacy need to be determined.

Elvira Famulari

Molecular Biotechnology Center, Italy

Title: Cell therapy for Crilger Najjar Type I syndrome

Time : 18:00-18:15

Biography:

Elvira Famulari is pursuing Ph.D in Molecular Biotechnology Center from University of Turin Italy.

Abstract:

Crigler-Najjar Syndrome type I (CNSI) is a recessive disorder caused by a rare mutation in Ugt1a1 gene that encodes for the enzyme Ugt1 involved in bilirubin metabolism. This mutation causes high levels of unconjugated bilirubin in blood, leading to brain damage and early lethality. CNSI patients are treated with extended daily phototherapy but the only effective therapy is liver transplantation. However liver transplantation is not exempt from complications, therefore hepatocyte-like cells derived from stem cells represent an interesting alternative. The goal of this project is to evaluate the use of human adult liver stem cells (HLSC) in treating Crigler-Najjar Syndrome type I (CNSI) in a model represented by Ugt1 deficient mice closely mimicking the pathological manifestations in CNSI patients. The Ugt1+/- mice have been backcrossed with NSG for 9 generations in order to derive these mice in a pure NSG background. By recellularising decellularised rat liver scaffolds with HLSCs ex vivo, we found that HLSC can differentiate in functional hepatocytes expressing Ugt1 protein as from 7 days differentiation. A significant improvement in survival of phototherapy-treated NSG/Ugt1a1-/- pups following injection of HLSC in the liver was observed. Immunohistochemically and immunofluorescence analyses show that HLSC can engraft in the liver of NSG/Ugt1a1-/- mice and express missing enzyme. HLSCs thus show great potential not only for the treatment of CNSI but also for most metabolic liver disorders.

Speaker
Biography:

Dr Wael Abo Elkheir has completed his Ph.D at the age of 35 years from Cairo University. He is the co-founder and board member of the Egyptian Society for Progenitor Stem Cell Research, a society initiated with the mission of enhancing scientific research and cooperation in the field of stem cell research and regenerative medicine. He is the director of a number of registered clinical trials in the field of stem cell therapy, especially for neuro-regeneration and musculoskeletal disorders. He has published more than 20 papers in reputed journals.

Abstract:

Background: Chondrogenesis is a well-orchestrated process in which chondroprogenitors undergo proliferation and chondrocyte differentiation. Because cartilage lacks blood supply, it shows poor regenerative power and subsequent wound healing. Cartilage degeneration caused by disease or trauma carries great clinical implication on the function of joints. The end stage of cartilage damage frequently leads to osteoarthritis (OA), resulting in a significant impairment of the quality of life of millions of people. MSCs are multilineage progenitors responsible for the normal turnover and repair of mesenchymal tissues, such as bone, cartilage, ligament, and fat. In-vitro expanded MSCs can differentiate into chondrogenic, adipogenic and
osteogenic lineages.
 
Aim: The objective of this work was to evaluate the regenerative effect of cell therapy in cases of acute and chronic OA canine models of surgically induced partial thickness chondral defects injected with autologous bone marrow derived MSCs.
 
Study Design: This work was done on 24 knees of male domestic mongrel dogs. OA was induced by doing surgical chondral defects then injected intra-articular with MSCs. Dogs were divided into the following groups: acute (injected after 1 day), chronic (after 1 month) and control group received conventional treatment. The dogs were sacrificed after 1, 2, 6 and 8 weeks
of injection. Assessment by histological scoring of cartilage repair (Os Score) for blind randomized samples and by clinical examination for lameness degree scores was done.
 
Results: Our results showed that dogs possess characteristics that are not found in traditional rodent models and confirmed the efficacy of direct intraarticular injection of MSCs to home and function in cartilage defects both in acute and chronic lesions.
 
Conclusion: This study concludes that the local delivery of MSC is a good therapeutic option for O A.

Speaker
Biography:

Hala Gabr is a renowned researcher in stem cell biology and therapy in Cairo University. She is the Director of the Pediatric Bone Marrow Transplantation and Cellular Therapy Lab in Cairo University. She is the Co-founder of the Egyptian Society for Progenitor Stem Cell Research, the leading stem cell research body in
Egypt. She has published more than 30 papers in reputed journals and is an Editorial Board Member of a number of reputed journals. She has supervised nearly a hundred Ph.D and Master thesis in stem cell research.

Abstract:

Liver transplantation represents the only definitive treatment for cases of end stage liver failure. However, this procedure is hindered by a number of obstacles, namely; the marked shortage of liver donors, major operative procedures and lifelong immune suppression, in addition to the high expenses. Regenerative medicine, based on cellular approach for repairing and replacing damaged tissues and organs, is a rapidly growing field of Medicine. Due to the heavy burden of liver diseases in Egypt, this study was designed to evaluate the efficacy of cellular therapy in the form of hepatocytes derived from patients' own haematopoietic stem cells (HSCs), transplanted directly through intra-splenic injections in patients with liver cirrhosis grade B and C Child-Turcotte-Pugh score (CTP B). 100 patients with liver cirrhosis CTP B score were divided into two groups according to the principle of treatment. Group (A) consisted of 50 patients (25 Child B and 25 Child C), who received hepatocytes derived from patients own (HSCs) in addition to conventional treatment. Group (B) received regular conventional treatment. Both groups of patients were followed up for six months after transplantation for assessment of liver functions. There was a significant improvement in the degrees of ascites, lower limb edema, HE, CTP scores and MELD scores in patients treated with hepatocytes derived from HSC. Also, we observed a slight improvement in serum albumin, prothrombin concentration and international normalized ratio in stem cell treated group. No procedure related complications were encountered. We demonstrated the safety and short term efficacy of autologous bone marrow derived hepatocyte transplantation for the support of cirrhotic liver.

Jin-Ye Wang

Shanghai Jiao Tong University, China

Title: Natural Polymer, Zein for Tissue Regeneration

Time : 17:00-17:20

Speaker
Biography:

Jin-Ye Wang has received her Ph.D from Tohoku University, Japan (1992). She was working as a Professor of Shanghai Institute of Organic Chemistry, Chinese Academy of Sciences (2000-2009), Adjunct Professor of Shanghai Jiao Tong University (2003-2009), and now Professor of Biomedical Engineering and Team Leader of Biomaterials Lab, Shanghai Jiao Tong University. She has published over 80 SCI papers, 4 books (chapters), and 13 Chinese patents and one US patent were authorized. She was the Invited Speaker of European Conference on Biomaterials, Pacifichem et al., awarded by the Hundred Talent Program of the Chinese Academy of Sciences (1999), Life Sciences Prize from Meiji Dairies Corporation (2008) et al. Her research interests include Tissue Engineering, Controlled Release and Fluorescent Probe, Biomimetic Materials and Biointerfaces
 

Abstract:

Zein is the major storage protein of corn and comprises 40-50% of total endosperm proteins. Zein has been used as microspheres to delay the release of drugs and to protect the drugs from degradation by pepsin, thus can release the
drugs for a long time. Our laboratory has developed zein as a novel and potential biomaterial for Tissue Engineering. Firstly, a three-dimensional zein porous scaffold was prepared and showed to be suitable for culture of various cell lines and primary cells such as human umbilical vein endothelial cells (HUVECs) and mesebchymal stem cells (MSCs) in vitro. The scaffolds are characterized with interconnected pore, controllable pore sizes, especially excellent mechanical properties, which are controllable and suitable to act as bone substitutes. Next, we examined its tissue compatibility in a rabbit subcutaneous implanting model, the histological analysis revealed a good tissue response and degradability. The third, zein porous scaffolds modified with fatty acids have shown great improvement in mechanical properties and also good cell compatibility in vitro. Besides, the complex of zein porous scaffold and mesenchymal stem cells (MSCs) could effectively promote the ectopic bone formation in nude mice and the repair of critical-sized bone defects in the rabbit model.

Shrikant L Kulkarni

Kulkarni Clinic, India

Title: Hydro-pressure therapy in chronic kidney diseases

Time : 16:40-17:00

Speaker
Biography:

Shrikant L Kulkarni has completed his MS (General Surgery) in 1975 from B J Medical College Pune, Maharashtra, India and his MBBS from Miraj Medical College,Sangli. Since 1971, he has worked at several Government Hospitals like the Wanless Hospital, Miraj, Sangli General Hospital, Sangli, Sassoon Hospital, Pune andMultispecialty Hospitals like Ruby Hall Clinic, Pune and Jehangir Nursing Home, Pune. For the last 35 plus years, he is working at his own clinic at Chinchwad,Pune, Maharashtra, India.

Abstract:

Self-organ regeneration or regenerative science is the body’s regenerative power, a natural physiological response of the organs or tissues when injured. Regenerative ability in the tissue awakens when required, indicating its proof of existence. Even today it is not clearly understood how cells recognize, what is missing or damaged and start regenerating these specific tissues. Self-repair mechanism would be the ideal solution for functional recovery of these failed organs. Today chronic kidney diseases (CKD) or end stage renal diseases (ESRD) patients are increasing day by day. Present treatment to these conditions is kidney organ allograft transplant or dialysis. The main cause for CKD is the presence of fibrosis in renal parenchyma producing toxic and hostile environment i.e. preventing the regenerative process. Fibrosis is body’s natural process, so dealing with this should be natural. Body can be repaired if fibrosis is dissolved to restore the circulation and elasticity of the arteries along with improving inflammatory immune system which makes the environment friendly in the damaged tissue.
 
The aim of this article is to discuss the use of the endogenous regenerative science for self-organ regeneration. A method based on hydro pressure therapy (artificially producing hydronephrosis) is discussed. Following steps are involved in this treatment) the pelvi–ureteric junction (PUJ) is blocked to create hydronephrotic condition, ii) due to increased back pressure the fibrosed renal parenchyma is dissolved iii) remove the artificial block at PUJ causing reduction in back pressure. Theoretically the prognosis, stem cells niches between renal capsule and the cortex will start regeneration of normal renal parenchyma. The hydro pressure therapy discussed in this paper, may prove an effective technique for renal replacement therapy which is safe, efficient and a low cost treatment.

Lin-Hwa Wan

National Cheng Kung University, Taiwan

Title: Application of ultrasonography in the assessment of overhand movement

Time : 16:20-16:40

Speaker
Biography:

Lin-Hwa Wang, a professor of the Institute of Physical Education, Health & Leisure Studies of National Cheng Kung University and the president of Taiwan Society of Biomechanics in Sports (2009-2011) in Taiwan, received the Ph.D. degree in Institute of Biomedical Engineering from National Cheng Kung University in Taiwan in 2010. She has been an outstanding researcher with high efficiency, precision and passion. Her work requires very little supervision and has earned high reputation in the areas of biomechanics. She has all of the attributes and potential that one might seek in a researcher. Main research focuses on relationship study on the biomechanical assessment of performance and injury of sports.

Abstract:

Statement of the Problem: Repetitive overhand movements generate significant mechanical stress on the shoulder and elbow joint. A specific focus on the ultrasound-based acute marker changes would make significant contribution to practical application to injury prevention, particularly to the design of intervention programs that reduce the likelihood of upper extremity injury. Through quantitative ultrasound examination of the elbow, acute changes in the joint structure at pre- and post-fatigue stages of similar overhand movements were investigated and the effects of fatigue on forearm muscle control over
overhand movements were determined.
 
Methodology & Theoretical Orientation: Ultrasonographical measurement of the length of the ulnar collateral ligament and its strain under the valgus stress test was performed. 12 tennis players performing first serves and 15 baseball pitchers performing fastball pitching were enrolled. The distances between the ulnar nerve and the tip of the medial epicondyle of the transverse images were measured in the various positions of the elbow, and a direct trace and ellipse tool provided by the highresolution ultrasonograph in this study helped the calculation of the distance.
 
Findings: The distance from the nerve to the medial condyle between the tennis players and baseball pitchers showed significant differences in the pre-fatigue stage (p = 0.006) as the elbow flexed to 90°.
 
Conclusion & Significance: Significant differences were observed in the distance from the ulnar nerve to the medial condyle between serving (tennis) and pitching (baseball) because the distance increases as the elbow flexes from 0° to 120°. Wide application of ultrasound imaging to investigate the impact of local fatigue on sports performance is expected in the future.

Speaker
Biography:

Dr. Yong-Jian Geng, in 1994, received doctoral degree in clinical chemistry from Goethenburg University, Sweden, and post-doctoral training at Brigham and Women’s Hospital, Harvard Medical School. 1997-1999, he joined faculty at Allegency General Hospital, Pittsburgh, and since University of Texas Health Science Center at Houston. He was appointed as director of Center for Cardiovacular Biology and Atherosclerosis Center and laboratory of Heart Failure and Stem Cell Research, Texas Heart Institute. He is a tenured, full professor of medicine, molecular pathology and bioengineering. His research work is focused on diagnosis, prevention and treatment of atherosclerosis-associated cardiovascular diseases as well as stem cell therapy.

Abstract:

Stem cell-based therapy has been emerging as novel treatment of various pathological conditions that routine medications are not effective to. Cardiovascular tissue regeneration and repair with different types of stem cells from adult tissues have been confirmed in both animal studies and clinical trials. However, autologous stem cell therapy has confronted a major obscle that stem cells from sick, aged patients with cardiovascular diseases are often malfunctional and have lower potency of regeneration and repair. To overcome this obscle, several approaches have been developed in this laboratory, including genetic or epigenetic rejuvenation, preconditioning with anti-atherogenic drugs, and enhancing survival and growth with cytokines and growth factors. Recent study has shown that transplantation of mesenchymal cells rejuvenated by overexpression of telomerase and myocardin, promotes revascularization and tissue repair in a murine model of ischemia. In vitro experiments further demonstrated that co-delivery of telomerase and myocardin genes rejuvenate adult stem cells to hightened myogenic development. Delivery of the iPS reprograming nuclear factor Oct3/4 improves the stem cell potential for tissue regeneration and repair. In atherosclerosis-prone mice, epigenetic modification of Oct3/4 gene promotor activity impacts the pathogenesis of atherosclerosis. Recent studies by this and other groups have documented evidence that treatment with statin, a cholesterollowering drup, augments the therapeutic efficacy of adult stem stem cells. Animal study has shown that atorvastatin enhances the efficacy of mesenchymal stem cells treatment for swine myocardial infarction via activation of nitric oxide synthase. Simvastatin-enhanced expression of promyogenic nuclear factors and cardiomyogenesis of murine embryonic stem cells.

Stephen S. Lin

California Institute for Regenerative Medicine, USA

Title: Initiatives to advance stem cell science and medicine at California’s $3 billion stem cell agency

Time : 15:20-15:40

Speaker
Biography:

Stephen S Lin is a Senior Science Officer at California’s stem cell agency, CIRM. He has joined the agency in 2015 to oversee the its $32M initiative to create a repository of iPSCs from over 2800 individuals covering both genetically complex and rare diseases, as well as a $40M genomics initiative that applies cutting edge genomics and bioinformatics approaches to stem cell research and therapeutic development. He is also Program Lead on a $15M award to create a preclinical research organization termed the translating center that focuses on preparing stem cell therapy candidates for clinical trials through support with process development, safety/toxicity studies, and manufacturing. From 2012 he had been a Staff Scientist and Team Lead at Thermo Fisher Scientific (formerly Life Technologies). Prior to that, he was a Scientist since 2006 at Stem Cells, Inc of California in liver cell therapeutics. He has received his Ph.D from Washington University in St. Louis in 2002 under Jeffrey Gordon and did his Postdoctoral research at Harvard University under Stanley Korsmeyer.

Abstract:

The mission of the California Institute for Regenerative Medicine (CIRM) is to accelerate stem cell treatments to patients with unmet medical needs. With $3 billion in funding and 300 active programs, it is the world’s largest institution dedicated to helping people by developing cell therapies. To accomplish its mission, CIRM has funded a breadth of activities spanning from basic research to translation to clinical trials. In addition to individual grants, CIRM has created resources to help the stem cell community worldwide. Some resources promote research and drug development using stem cells. CIRM has established an iPSC Repository maintained by the Coriell Institute that is currently the largest publically accessible pluripotent stem cell bank in the world. CIRM also has a genomics research initiative that applies cutting edge sequencing and bioinformatics approaches to stem cell research and therapeutic development. Other resources established at CIRM promote the acceleration of stem cell discoveries into therapeutic development, including the Stem Cell Center, which supports preclinical IND-enabling and clinical trial management activities related for stem cell therapies, and an Alpha Clinics network that conducts clinical trials for stem cell related therapies. CIRM supports additional activities that promote standardization, clarity, and speed to this emerging therapeutic area. In total, these resources are designed to catalyze the flow of stem cell discoveries to the clinic, which can come from both inside and outside of California.

Speaker
Biography:

Seungil Ro has obtained his Ph.D in Cell and Molecular Biology from the University of Nevada, Reno, USA in 2002, where he has been Associate Professor since 2015 in the Department of Physiology and Cell Biology in School of Medicine. His research interest includes the roles of microRNAs that regulate gastrointestinal smooth muscle motility and epigenetic remodeling. He has 35 papers published to his credit.

Abstract:

Transcriptome data on the quantitative numbers of transcriptional variants, expressed in primary cells offer essential clues in cellular functions and biological processes. We obtained transcriptomes from intestinal primary cells (smooth muscle cells, SMC; interstitial cells of Cajal, ICC; PDGFRα+ cells, fibroblast-like cells) and tissues (jejunal and colonic smooth muscle and mucosa). We built “Smooth Muscle Transcriptome Browser” and “Smooth Muscle Genome Browser” that can offer genetic references and expression profiles of all transcripts expressed in SMC, ICC, PDGFRα+ cells, associated jejunal and colonic tissues. Using these browsers, analyzing the transcriptomics, we have identified a unique set of cell signature genes for the
three cell types including growth factors, transcription factors, epigenetic enzymes/regulators, protein kinases/phosphatases, cytokines/chemokines, receptors, and ion channels/transporters. We found that the cell signature genes are dysregulated in many gastrointestinal diseases and can serve as new pathological markers and therapeutic targets. Taken together, Smooth Muscle Transcriptome Browser and Smooth Muscle Genome Browser bring new insights into the cellular and biological functions of primary SMC, ICC, and PDGFRα+ cells in gastrointestinal smooth muscle biology and diseases.

Speaker
Biography:

Joanne Mullarkey has been an Adult Nurse for 11 years, working initially on ICU and HDU wards in a large teaching hospital in Leeds UK. She has also worked in
a private hospital for 5 years and started working for a Human Tissue Bank based on the University of Bradford city campus. She has developed the TDAD initiate
over the past 3 years and looking to expand the service even further in the next 2-5 years. She is GCP trained and an experienced Communicator.

Abstract:

Ethical tissue has been designed to give researchers access to quality tissue samples, optimized usage of donations and provision of linked clinical data. There are almost 300 licensed tissue banks in the UK; however, Ethical tissue is unique in the way it can supply tissues to industry and academia. Ethical tissue is distinctive in collecting any type of tissue on a bespoke basis, providing it directly to pre-approved researchers all over the world. A key issue for many researchers has been their need for bespoke collections of tissue that are impossible to obtain via surgical intervention. From this, we identified the need for a Tissue Donation after Death (TDAD) program. Donors who cannot donate an organ/tissue for therapeutic can use TDAD to
support biomedical research. Feedback from donor families indicates how much satisfaction donors derive from being able to contribute in this way. From its inception in 2014, we have developed the TDAD service from an initial single donation to the point where we expect to have at least 30 donors this year. TDAD now forms a key part of our tissue provision, particularly for
difficult to obtain tissues. In building up the TDAD service, a lot of time and energy has gone into educating local healthcare professionals and the general public on tissue donation and end of life choices which are available to them. It does however require changes in thinking and working practices, not always easy particularly within large organizations. What has been achieved through TDAD and our other services has culminated in ethical tissue being recognized as the UKCRC Biobank of the Year (2016/17).

Biography:

Vittorio Sebastiano is a Stem Cell and Developmental Biology Expert. His research focuses on dissecting the mechanisms that regulate nuclear reprogramming with the goal to use patient specific stem cells for the development of clinically relevant platforms for the treatment and the modeling of pediatric orphan diseases. He has been among the pioneers in gene therapy by in situ specific genome modification of iPSC derived from patients affected by Sickle Cell Anemia and Epidermolysis Bullosa. Currently, he is utilizing iPSCs to model developmental disorders and he is implementing clinical platforms to develop cell therapeutics for Precision Medicine.

Abstract:

The discovery of methods to convert somatic cells into induced pluripotent stem cells (iPSC) is probably one of the most revolutionizing breakthroughs in Regenerative Medicine within the last decade. iPSC are functionally equivalent to Embryonic Stem Cells (ESC) and have the ability, defined as Pluripotency, to generate any cell type of the adult body. Unlike ESC though, iPSC can be derived from the tissues of any individual, raising the possibility of producing custom-tailored cells for the study and treatment of virtually any disease. Furthermore, they are amenable to genetic manipulations, including
homologous recombination (HR), which allows the in situ correction of the disease-causing mutation and avoids several safety risks associated with conventional vector-based gene therapy involving random integration such as non physiological gene expression and cancer formation. Although these prospects are exciting, several hurdles are associated with iPSC technology.
Questions arise about the safety of the reprogramming and gene targeting methodologies, which involve extended culture periods, differentiation efficiency, and quality of iPSC-derived cells. These questions need to be answered before translation of iPSC-based technologies to the clinic. Research in the Sebastiano lab focuses on all aspects of this technology, spanning
from nuclear reprogramming, to genome editing and in vitro differentiation. The ultimate goal of the lab is to develop clinical therapeutics using patients-specific iPSCs, by both modeling genetic and developmental disorders in vitro and by developing stem cell therapies that will be soon utilized in the clinic. Proof of principle studies and data on Epidermolysis Bullosa and DiGeorge Syndrome will be presented and discussed.

Sita Somara

Wake Forest Institute for Regenerative Medicine, USA

Title: Clinical translation of tissue-engineered medical products (TEMP) - Journey from bench to bed-side

Time : 14:00-14:20

Speaker
Biography:

Sita Somara has completed her Ph.D from SK University, India and Post-doctoral studies from University of Michigan, Ann Arbor. She was appointed as Assistant Professor at Wake Forest Institute for Regenerative Medicine in 2011. She has been served as Investigator in NIH-funded and private foundation projects. In 2014, she joined Regenerative Medicine Clinical Center as Lead Process Development Scientist to pursue her passion for Translational Medicine. She has since led and managed TEMP project in highly regulated environment developing cell-based products, tissue-engineered products and cell/tissue banking in GLP and GMP. She has authored FDA submissions from PPIND, PIND and IND specifically CMC sections. She has published more than 25 peer-reviewd publications in reputed journals. She has recently been appointed as a Committee Member of “Legal and Regulatory Affairs Committee: North America” of International Society of Cellular Therapy (ISCT).

Abstract:

Tissue-engineered medical products (TEMPs) are rapidly growing as an advanced innovative therapeutics that can restore, repair and regenerate the cells and tissues to improve functionality. The driving force behind the rapid growth of TEMPs is unmeet clinical need of organs and organ donors. Tissue engineering uses a novel interplay of cell and biomaterial and very efficiently brings life science and engineering concepts together. Presentation will illustrate process development of translating TEMP to clinical manufacturing with focus on regulatory requirements with a case study. Preclinical safety studies involving both in vitro and in vivo using small and large animal models that help find solutions to key research questions will be discussed. Ethical issues in clinical studies with regards to use of cells and tissues, their sources, donor consent, as well as clinical trials will be addressed. TEMPs are regulated as drugs, biologics, devices, or combination products by the US Food and Drug Administration (FDA). Institutional and government levels regulatory issues must be addressed prior to the translation of TEMPs to clinic. The presentation will highlight the regulatory issues for tissue engineered medical products that assures quality, safety and efficacy. Presentaion will also discuss how Regenerative Medicine Advance Therapy (RMAT) also called 21st Century Cures Act will help fasten the translation process of TEMP.

Speaker
Biography:

Bo Feng is an Assistant Professor at the School of Biomedical Sciences, Faculty of Medicine, The Chinese University of Hong Kong. She is an active Staff Member in the Stem Cell and Regeneration program, MOE key laboratory for Regenerative Medicine and CUHK-GIBH joint laboratory on Stem Cell and Regenerative Medicine. She graduated from Nankai University with BSc (1993) and MSc (1996), and received her Ph.D (2006) from National University of Singapore. After graduation, she joined Prof. Ng Huck Hui’s lab in Genome Institute of Singapore as a Post-doc. She worked on stem cells and reprogramming and published her works in Nature Cell Biology, Cell Stem Cell and Nature. In Nov 2010, she joined CUHK and her current research interest lies within the molecular mechanism that controls pluripotency and differentiation of ESCs/iPSCs, as well as development of new tools for stem cell research and applications.

Abstract:

CRISPR/Cas9-induced site-specific DNA double-strand breaks (DSBs) can be repaired by homology-directed repair (HDR) or non-homologous end joining (NHEJ) pathways. Extensive efforts have been made to knock-in exogenous DNA to a selected genomic locus in human cells; which, however, has focused on HDR-based strategies and was proven inefficient. Here, we report that NHEJ pathway mediates efficient rejoining of genome and plasmids following CRISPR/Cas9-induced DNA DSBs, and promotes high-efficiency DNA integration in various human cell types. With this homology independent knock-in strategy, integration of a 4.6 kb promoterless ires-eGFP fragment into the GAPDH locus yielded up to 20% GFP+ cells in somatic LO2 cells, and 1.70% GFP+ cells in human embryonic stem cells (ESCs). Quantitative comparison further demonstrated that the NHEJ-based knock-in is more efficient than HDR-mediated gene targeting in all human cell types
examined. These data support that CRISPR/Cas9-induced NHEJ provides a valuable new path for efficient genome editing in human ESCs and somatic cells.

Pao Chi Liao

National Cheng Kung University, Taiwan

Title: Biomarker discovery using mass spectrometry-based proteomics/metabolomics and biospecimens

Time : 12:15-12:35

Speaker
Biography:

The introduction of sensitive ionization methods and continuing improvement in resolving power in the past decades has made mass spectrometry an ideal tool for biomarker discovery. The coupling of chromatography and mass spectrometry emerged to be the most powerful profiling techniques for global characterization of proteins and metabolites in biological systems. In this “omics” era, many researchers rely on mass spectrometry-based proteomic and metabolomics experimental approaches to search for potential biomarkers. In this talk, I will describe how I applied these approaches to (1) discover metastasis-promoting secretory proteins of lung cancer cells; (2) discover virulence factors secreted from Streptococcus pyogenes in response to wound environments; and (3) discover biomarkers for assessing exposure to toxicants. Challenges and limitations will be discussed.

Abstract:

Dr. Pao-Chi Liao completed his Ph.D. in Analytical Chemistry from Michigan State University (MSU) in 1995 before doing postdoctoral research in the Department of Biochemistry at MSU. Dr. Liao joined the faculty at Department of Environmental and Occupational Health, National Cheng-Kung University, Taiwan in 1997, where he was promoted to full professor in 2006, and named Distinguished Professor in 2011. Dr. Liao’s research interests and fields of specialty include analytical chemistry, mass spectrometry, proteomics, biomarker discovery, cancer biomarkers, lung cancer metastasis, and environmental and occupational health.

Speaker
Biography:

Won-Gun Koh received his BS and MS degrees from the Department of Chemical Engineering at Yonsei University, Korea. He received Ph.D in 2004 from Department of Chemical Engineering at Penn State University under the guidance of Professor Michael V Pishko. After Ph.D, he was appointed as Post-doc scholar at Stanford University, where he made relationship with Professor Curtis W Frank. At Stanford University, he worked in the Artificial Cornea Project with collaboration of Stanford University School of Medicine. He became the Assistant Professor in the Department of Chemical and Biomolecular Engineering at Yonsei University in 2005. Currently, he is Professor in the same department and Director of Yonsei Center for Research Facilities. His research interests include polymerbased tissue engineering and biosensor.

Abstract:

In this study, we describe a simple method for fabricating multiscale scaffolds that are capable of controlling the spatial positioning of mammalian cells and proteins or peptides. Photopatterning of poly (ethylene glycol) (PEG) hydrogel on the electrospun nanofibers produced micropatterned nanofiber matrices made of hydrogel microwells filled with a nanofibrous region, which is capable of generating cell and protein micropatterns due to the different interactions that cells and proteins have with PEG hydrogels and nanofibers. Different proteins could be immobilized onto resultant micropatterned nanofiber scaffold, carrying out cell patterning, metabolite detection, and growth factor delivery. As potential applications of resultant scaffold, the control of stem cell differentiation via controlled release of multiple growth factors. We fabricated fibrous scaffolds incorporating PEG hydrogel micro patterns for potential applications to spatio-temporal release of multiple growth factors for optimized osteogenesis of hMSCs. The resulting scaffolds were capable of loading different growth factors separately into the hydrogel micro patterns and the fibers in a single scaffold platform. Sequential delivery of bFGF and BMP-2 for osteogenesis of hMSC was achieved by a quick release of bFGF from the fibers and a slow and sustained release of BMP-2 from PEG hydrogel. The enhanced effect of the sequential release of bFGF and BMP-2 on the osteogenesis of hMSCs was clearly validated by in vitro
studies such as ALP activity/staining and mineralization studies.

Speaker
Biography:

Nidhi Bhutani is an Assistant Professor in the Department of Orthopaedic Surgery at Stanford University and is affiliated with the Cancer Biology Program, the BioX program and the Children Health Research Institute at Stanford. Her research interests broadly encompass the molecular mechanisms regulating regeneration and repair of the musculoskeleton, with a focus on epigenetic regulation by DNA methylation and demethylation. Her group is interested in applying stem cell and reprogramming based approaches towards musculoskeletal tissue engineering.

Abstract:

Regeneration of human cartilage is inherently inefficient, a key factor for the widespread occurrence of degenerative diseases like Osteoarthritis (OA). Recent reports have provided compelling evidence that juvenile chondrocytes (from donors below 13 years of age) are more efficient at generating articular cartilage as compared to adult chondrocytes. However, the molecular basis for such a superior regenerative capability is not understood. We aimed to identify the cell-intrinsic differences between young and old cartilage and systematically profiled global gene expression changes between a small cohort of human neonatal/juvenile and adult chondrocytes. Our studies identified and validated new factors enriched in juvenile chondrocytes as compared to adult chondrocytes including secreted ECM factors Chordin-like 1 (CHRDL1) and Microfibrillar-associated protein 4 (MFAP4). CHRDL1 was observed to aid the proliferation and survival of human bone-marrow derived mesenchymal
stem cells (MSC) providing a mechanism for how young cartilage factors can potentially enhance stem cell function in cartilage repair. Similarly, we observed that soluble Collagen VI (Col VI) enhances the proliferation of adult chondrocytes without any dedifferentiation. These juvenile cartilage factors are therefore useful in maintaining chondrocytes and can be potential biologics with useful applications towards biobanking of mesenchymal stem cells, chondrocytes and cartilage constructs.

Jill Davies

Oxford University, UK

Title: Challenges of developing a biobank in Oxford, UK

Time : 10:35-10:55

Speaker
Biography:

Jill Davies graduated from Coventry University in 1987 with a degree in Applied Biology. After university studies, she worked for cardiothoracic surgeons Mr. Donald Ross & Sir Magdi Yacoub at the National Heart Hospital in London and that she opened the heart valve bank at the Oxford University Hospital in Oxford, England in 1990. The Oxford bank supplies cardiovascular tissue, corneas for transplant and research and brains & spinal cords for research. It is now also an Oxford Research Center Bio bank. She is also an executive member of the BATB, member of AATB, SLTB and Society for Cryobiology.

Abstract:

Oxford University Hospital Cell and Tissue Biobank (OCTB) now provides a range of tissues for research purposes e.g., fertility tissue from patients undergoing sterilizing cancer treatment and neurological tissue from deceased patients. This development into bio banking domain has posed new challenges. Very few people in UK register as donors who exasperate coordinators asking emotional cancer patients/parents or bereaved families for consent (consent is therefore recorded). Areas of public concern are discussed (e.g., use of tissue in animal studies) and these options are therefore more often declined. Oxford re-consent donors when they become 18. Oxford has also overcome issue of collection of brain/spinal cord from patients who don’t have mental capacity to consent. OCTB is within UK National Health Service (not-for-profit) therefore direct/indirect payment to donors is unacceptable. Payments for use of facility (mortuary/theatres) or staff performing tissue
retrievals is problematical. Reimbursement of costs to biobank following tissue release is difficult to estimate and may not be affordable to researchers. Agreements set up between tissue bank and researchers to confirm ethical, research study approval, sponsors restrictions. Publication acknowledgement, patent ownership and secrecy issues with private funded projects pose
difficulties (UK competent authority must review validation documentation). Legal, quality and safety framework is enforced across Europe but harmonization of operational/ethical issues is ongoing. Oxford has comprehensive quality management system and undergoes annual inspection and licensing. Tissues are directly distributed by OCTB without patient identifiable details. Feedback is not given to donors/bereaved families, even if something medically beneficial is established. All tissues are traceable from donor to end user using single European code and bar coding. Marketing of tissues/cells is authorized in theUK. Oxford is registered in UK tissue biobank directory without a price list. These details could decrease goodwill of donors, increase bad publicity, encourage nefarious trading. Commercial distributors are licensed in the UK, numbers increase as demand increases. General public not yet alerted to this trading. Final costs of tissues released are not controlled or capped in the UK. There are no schemes to monitor/measure needs of researchers. Oxford only permits export if donor was offered this option during consent interview. The Oxford multidisciplinary teams meet regularly to discuss ethical and operational issues. Service review with patients/parents and bereaved relatives is invaluable. This approach has enabled the rapid development of OCTB biobanking service.

Jan Huebinger

Max-Planck Institute of Molecular Physiology, Germany

Title: Cryopreservation of living cells using electron microscopy fixation methods
Speaker
Biography:

Jan Huebinger has his expertise in the development of cryo-immobilization techniques of living samples. He co-developed and evaluated new methods for cryofixation for cryo-electron microscopy and for cryopreservation of living cells. More recently he has co-developed a method that allows to reversibly cryo-arrest living cells on a light microscope. This allows to image highly dynamic processes in living cells with methods that necessitate long acquisition times, e.g., super resolution microscopy, at multiple points in time.

Abstract:

Rapid cooling to minimize ice crystal growth can be applied for cryopreservation as well as sample preparation for (electron) microscopy. However, the methods used in these two fields are very different. To preserve the structure of living samples for electron microscopy in a close to native state, they have to be completely vitrified with minimal use of cryoprotective
agents. We found that complete vitrification is not necessary for successful cryopreservation of mammalian cells. However, ice crystal size (not number or total amount of ice) has to be minimized. Therefore, sample preparation methods for (electron) microscopy could also help improving the outcome of cryopreservation. We found that standard preparation methods are not very suitable for cryopreservation, likely because rapid warming is not possible with these methods. However, we found that the recently developed method of self-pressurized rapid freezing is very suitable for cryopreservation. Sealed metal tubes with high thermal diffusivity containing the samples are plunged into liquid cryogen. Internal pressure builds up reducing ice crystal formation and therefore supports reversible cryopreservation through vitrification of cells. After rapid rewarming of pressurized samples, viability rates of >90% can be reached, using human cells (HeLa). This is comparable to best-performing of the established rapid cooling devices tested. In addition, the small SPRF tubes allow for space-saving sample storage and the sealed containers prevent contamination from or into the cryogen during freezing, storage, or thawing.