2^nd International Conference & Exhibition on Tissue preservation and Bio-banking September 12-13, 2016 Philadelphia, USA

Biography:

Mohamed A Zayed is a Surgeon-Scientist at Washington University School of Medicine, Department of Surgery and Section of Vascular Surgery. He has completed his medical training at Stanford University, and Doctoral degree in Pharmacology at the University of North Carolina at Chapel Hill. He has served as Chief Medical Officer for a software start-up company, and has published over 25 research articles in reputed journals. His current clinical and research interests focus on “The influences of diabetes on peripheral arterial disease”. This field provides a unique appreciation of the complexity and potential gaps of knowledge in vascular pathophysiology and its end-stage complications.

Abstract:

Objectives: Over three 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 22 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: 356 patients were enrolled in the vascular surgery biobank prior to the index operation. 42 clinical variables were evaluated for each patient during the perioperative period. Vascular specimens were successfully collected for 54% (193) of patients who enrolled in the biobank. The majority of specimens collected were retrieved from the peripheral arterial system (50% carotid artery, 15% anterior or posterior tibial arteries, and 12% femoral or popliteal arteries). Each arterial specimen was sub-divided into maximally and minimally diseased portions to facilitate intra- and inter-patient biochemical and molecular analyses.

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, and subsequent experimental analysis.

Biography:

Arredondo E (BS, MSc, TPM) is a Biologist specialized in Organ, Tissues and Cell Donation from the University of Barcelona. He has participated in the implementation of liver tissue for hepatocytes isolation and SELICA clinical trials. He is currently responsible for the tissue at Research Department of DTI Foundation. His studies are based on how to develop networks to procure human tissue for research. He has been involved in several European projects related to organ donation such as the BSA project sponsored by the Council of Europe.

Abstract:

Introduction: The human hepatocytes in vitro model are the gold standard regarding a wide range of applications in biomedical research such as biological, pharmacological and toxicological studies. Researchers face many ethical and legal challenges in this area, and also sourcing high quality, fully characterized specimens able for use in bioanalytical processes.

Objectives: Aim of this study is to provide fresh human hepatic tissue fully characterized with high quality and viability criteria for hepatocytes isolation for research purposes.

Materials & Methods: The tissue was procured from patients undergoing planned liver resection surgeries due to primary or secondary tumors (living donors). The program required the involvement of health professionals in donor detection, tissue retrieval and evaluation, tissue perfusion and isolation. Hepatocytes isolation was conducted through the two-step collagenase perfusion technique and the established in-house methods. Cell viability and yield production were assessed and linked to donor clinical information. The program follows the European legislation to ensure quality, safety and traceability of all the procedures.

Results: The program linked eight procurement centers. A total of 222 donation offers were reported and evaluated; 116 donations have been isolated under a cold ischemia time of 8 hours. The 86.66% of the isolated tissues produced viability >80% and an average of 16.49 million of hepatocytes/gram were obtained.

Conclusions: Thanks to a consolidated hospital network for providing fresh tissue for research with high quality and viability criteria. As a result of the program, studies of the influence of donors’ pre-, intra- and post-operative parameters have been performed.

Biography:

Veronica Ponce de Leon has received her PhD in Biochemistry at the École Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland, in 2009 and further specialized in Genetics and New Genetic Engineering Technologies. In parallel to her scientific career, she moved on to start her own company: Innovation Therapeutics, to cure eye diseases through Ex-vivo Cell Therapy. In 2014, she won an InnoPACCT grant and the Isabelle Musy Prize for Women Entrepreneurs. She has joined Genohm as Vice President of the US branch Genohm Inc., to lead the development of its business in the States

Abstract:

Modern biobanking must be easy to manage with high standards of quality control. High-quality repositories acquire, accumulate and consolidate data from various sources and in various formats to build their knowledge base over time. The LIMS+ELN+order management features of SLims are designed to deliver a seamless management and processing of bio-specimen and samples of the research community. SLims is a flexible and compliant connectivity platform that supports and links different software, computers, barcode readers, label printers and all other relevant instruments of your lab and biobank. With SLims you are able to register, label, store and manage patients’ samples and easily link them with data, protocols and results. Integration of new content and patient’s follow-up is seamless. The main key features of the SLims platform for biobanking are: Quick and easy overview of sample entries, storage locations, sample status, sample workflow; Highly configurable sample annotation with first-rate label templates and printing options; Genetic sample relations with pedigree information (family relations); Quick standard operations, configurable workflows and easy order processing; Role-based data validation options; Full scale chain of custody, perfect for research project management (RUO settings); Controlled and selective access, 21CFR 11 compliant; Sophisticated query options, customized exports and reports; Integration possibilities for freezer management systems, instruments and hospital systems; Highly adaptable environment, ready for the future.

Biography:

Paul Murphy is a Cryopreservation Sales Specialist of Hamilton Storage, a worldwide provider of automated sample storage systems with headquarters in Franklin, Switzerland.

Abstract:

Hamilton Storage provides ultra-low temperature automated sample management solutions for the life science industries. Products include Askion C-line® systems for -150ºC cryopreservation, Hamilton BiOS® and SAM™ for -80ºC biobanking and Verso™ and ASM™ for -20ºC high-throughput tube and plate management systems. Our new consumables, RackWare™, are high-density SBSfootprint racks for tubes and cryovials designed to increase the storage capacity in our BiOS, SAM and Verso systems. The LabElite® line of benchtop devices includes an automated decapper, automated 2-D barcode reader and a combination of the two devices to provide flexible solutions for sample processing, tracking and security. Furthermore, our systems integrate with Hamilton Robotics’ automated liquid handling workstations for complete biobanking solutions.

Biography:

Elena Salvaterra is a Jurist Doctor (JD) and PhD in Law of Science (Bioethics). After a few years spent at academia (University of Milan), she collaborated with the Biological Resource Center of the Polyclinic University Hospital in Milan as Social Scientist. She has co-founded with Dr. Paolo Rebulla, Hematologist, the Laboratory of Bio-Law, dedicated to the study of ethical, regulatory, economic and social aspects associated with therapeutic and research human biobanks. She is presently a Consultant for Biobanks at Air Liquide Healthcare in Milan (Italy) and is a member of the Scientific Advisory Board of CloudLIMS. Her research areas cover Biobanks ELSI, economics, neuroethics and neurolaw, robot-ethics, ethical and regulatory issues of medical assisted procreation and ethics of euthanasia. She has recently edited the book “Regulating biobanks in humans: The use of adult and children biomaterials for clinical and research purposes”. 

Abstract:

Emergencies can occur in biobanking life every time with relevant impact on business continuity. While disaster recovery plans are crucial for biobanking sustainability, they are not usually considered or properly written for large as well as for medium and small biobanks. This presentation aims to give an overview on disaster recovery plans for biorepositories by focusing on: Regulatory requirements, technical procedures and organizational aspects. Case studies on happened disasters will be summarized by taking a picture of biobanking emergencies worldwide. Basic instruction about how to write a plan for disaster prevention, mitigation and recovery; and how to put in place it will be provided. 

Biography:

Dr. Mitchel C. Schiewe attained a BS/MS at UC Davis (1981)/LSU (1983) focused on Animal Reproductive Physiology. Working with the Smithsonian Institution/National Zoological Park and the NIH, he completed his PhD in Human Physiology in 1989 at the Uniformed Services University of the Health Sciences (Bethesda, MD). Subsequently, he performed his post-doctoral studies at NIH/NCRR as an NSF Associate. He is currently a Scientific/Technical Lab Director for Ovagen Fertility and the California Cryobank. He considers himself to be a Comparative Reproductive Physiologist, specializing in Embryology, and has published more than 35 peer-reviewed papers and 110 scientific abstracts, including several research award presentations.

Abstract:

Biography:

Henry QX Li, Ph.D. currently leads Crown’s Translational Oncology Division with platforms built on patient derived xenograft (PDX). Prior to Crown, Dr. Li has over 20 years of biopharmaceutical, as well as academic R&D, experiences in cancer and viral infection, including leadership roles as R&D director/senior director roles in several US-based biotech companies. Dr. Li currently also holds visiting Professor position at Peking University-State Key Laboratory. He earned his Ph.D. in Molecular Biology/Biochemistry from University of California (Irvine) and completed his postdoctoral training at UCLA School of Medicine. He has published more than 50 manuscripts and edited 3 books in the biopharmaceutical areas. He is also on the editorial board of Current Signal Transduction Therapy.

Abstract:

Cancers are diverse diseases, with various histo-/molecular pathogenesis, and respond vastly different to treatments. Optimal therapy needs to be precise and personalized, which demands accurate diagnosis and modeling. We recently developed a new molecular diagnostic tool, HuDiagnosis^TM, that can precisely diagnose cancer diseases using NGS per a novel molecular pathology algorithm, to guide precision therapy.  This tool, with equivalency to classic histopathology but significantly more reliable and objective, can also be particularly useful in annotating biobank.  In addition, we have also built the largest patient derived xenograft (PDX) library of >3,000 models covering divers cancer types with full annotation of histo-/molecular pathology, growth and treatment information.  This growing library, with intent as experimental model to support population-based investigations (clinic styled) at preclinical setting (mouse clinical trial -- MCT), is also a live and renewable biobank that can be used to facilitate various biobank functions, e.g. biomarker discovery.  In particular, together with experimental efficacy readout, this live bank can greatly enhance predictive biomarker exploration.   

Biography:

Mary A Hall completed her PhD from Southern Illinois University at Carbondale in Molecular Biology, Microbiology, and Biochemistry, with a focus in Neuroendocrine Immunology; and MBA from the University of Houston, with focus in business administration, management, and marketing. Her Post-doctoral research was conducted at Research Inc., Veterans Affairs Medical Center in Memphis, TN; and the Department of Immunology, MD Anderson Cancer Center in Houston, TX. He is the Program Manager at the Center for Clinical and Translational Sciences (CCTS) Biobank at The University of Texas Health Science Center at Houston (UTHealth). 

Abstract:

During the past 20 years, sample and data repository evolution has contributed to rapid biomedical research growth. Much effort has been directed toward customizing information and communication technology platforms to integrate and make available normalized clinical and research data for big data sharing among researchers to advance discoveries and translation to clinical practice. Here, we adapted the Informatics for Integrating Biology and the Bedside (i2b2) platform to manage clinical, genomic, and other study participant-related data for a biorepository at The University of Texas Health Science Center at Houston (UTHealth) to facilitate collaborative sharing of samples and data by researchers. We developed our customized Sample Location and Enhanced Distribution (SLED) application to run on i2b2 open source code. Clinical diagnoses, demographics, medical histories, sample types, and other de-identified data from 10,221 consenting study participants were imported into SLED, which researchers now use to query data and submit requests for samples and data to the UT Health Center for Clinical and Translational Sciences (CCTS) Biobank. Query results are relayed to researchers within seconds and submitted requests are delivered immediately to the Biobank. Since SLED production implementation, we have documented 156 logins by 25 distinct users, with 81 queries run, and 6 requests for samples or data submitted via SLED. In summary, SLED provides automated management of integrated clinical and research data at the UTHealth CCTS Biobank, including researcher queries and requests. The i2b2-based SLED design may be suitable for others seeking to customize a scalable and portable application for managing biorepository data. 

Biography:

John M Baust (President of CPSI Biotech) is a recognized Innovator in Cryomedicine. His research has been instrumental in the advancement of the cryobiological sciences into the molecular era and is credited with the discovery of cryopreservation-induced delayed-onset cell death. He has published more than 100 peer-reviewed papers, reviews and chapters. Currently, he serves the Editorial Board of Biopreservation and Biobanking, Board of American College of Cryosurgery. He has completed his studies at Cornell University, Binghamton University and Harvard Medical School.

Abstract:

Cryopreservation (CP) plays an integral role in a variety of bioprocessing, biotechnology and medical applications. While a critical tool, CP protocols, approaches and technologies have evolved little over the last several decades. While the adoption of new approaches to CP has been slow, discoveries including molecular modulation and the development of new devices for improved sample freezing and thawing are providing new strategies for improving CP. To this end, we have developed a series of new devices and protocols to enable the rapid and controlled freezing (Smart Freeze) and thawing (SmartThaw). These systems are designed to improve sample viability and function post-thaw while reducing processing time and end-user variability. This presentation will discuss these new systems as well as the impact of molecular stress response and the apoptotic process on CP outcome. Data presented will include thermal profile, cell viability and molecular stress results from several cell systems including CHO, PC3, HUVEC and hMSCs. The results suggest that these systems enable more efficient, controllable sample processing in comparison to traditional methodologies. Importantly, these investigations are providing new technologies and directions, built on a cell/molecular foundation to help accelerate new research, technology and procedure development initiatives in which CP serves as an enabling component.

Biography:

After many years of wet lab experience and living in the United States, Charles Wang decided it was time for a change of scenery and moved back to China in 2012. He is recruited specifically by his professional skill set and experience in scientific solutions for biobank and informatics. He has earned his leading role in the young field of biobanking informatics in China. He is traveling between China and USA for his new challenges and promotes international collaboration potentials. He is not only recognized for his dedication to biobanking, but also to data harmonization and sharing model development.

Abstract:

Data sharing across cohort studies for joint analysis is the trend of etiology study by a large number of cases. Shanghai Birth Cohort (SBC for short) banks samples collected from preconceptional care, which links to the questionnaire data and medical records. Two birth cohorts by Canadian team share many key elements of scientific questions with SBC. This is thus a great opportunity for the teams to share the resources. However, study design and information collection vary with studies, which generate data heterogeneity across the studies, not speaking of ethical and legal barriers that also challenge data sharing and international collaboration. By the joint effort, the two teams have launched a collaboration aiming to capitalize on data harmonization to pave the way of data sharing. The approach for data harmonization begins with the datasets which are selected based on mutual research interests and harmonization potentials. With the harmonized datasets, we apply two different approaches for joint analysis: We take “federated analyses” approach in which the joint analysis is conducted with DataSHIELD technology, which allows to share descriptive analysis across cohorts without sharing individual-level data. This approach is to bypass ethical and legal restriction across cohorts and countries to facilitate international collaboration and; the team has strong interest for further collaborative study, we thus take “pooled analysis” approach, which allows to pool the data for further analysis under the established ethical framework for good ethical governance. In a word, harmonization of datasets is essential for biosample and data sharing.

Biography:

Xiaopu Yuan has completed his MD in 2^nd Medical University in Shanghai China. He had been practicing surgical pathology for 13 years before immigrate to US. After pathology fellowship program in Mayor Clinic, he joined FHCRC as a Research Pathologist and worked there for 12 years. Since 2011, he has been working in Biobank and Translational Research Core at Cedars-Sinai Medical Center as a Research Pathologist. He has publications in China and US.

Abstract:

To provide researchers with the highest quality prostate cancer tissues, we developed a new protocol for tissue collection from prostatectomies. Since prostate cancers cannot be grossly appreciated, microscopy is required to identify tumor areas. We collected 2,737 frozen blocks from 415 consented patients in 2010-2014 under an IRB-approved protocol. During regular submission of radical prostatectomy cases in the clinical pathology laboratory, two central slices of the prostate were obtained for banking. The inked margins were removed and with the rest of the specimen. The remainder was frozen into up to eight cassettes. After the case was signed out, a diagnosis was obtained for all frozen blocks. Two anatomic pathologists reviewed the slides and recorded the tissue size, tumor size and percentage, Gleason sum (GS) and percent inflammation. 451 out of 2,737 blocks contained cancer (16.5%) while 174 out of 415 cases contained cancer (41.9%). The greatest diameter of the tumor ranged from 0.1-0.3 (22%), 0.4-0.9 cm (48%) and ≥1cm (30%). GS 6 was present in 24.8%, GS 7 in 46.1%, GS 8 in 10%, GS 9 in 16%, and GS 10 in 3.1%. Seventy slides contained a tertiary score (15.6%). Inflammation amounted to 10% in 56.1% of slides, 20% in 35.7% of slides and 30-40% in 8.3% of slides. For each slide, histological parameters were entered into designated and searchable fields within the OnCore database. Altogether, our protocol has created a precise inventory in the biobank and significantly increased the speed and efficiency of distributing prostate cancer tissues.

Biography:

Mohamed A Zayed is a Surgeon-Scientist at Washington University School of Medicine, Department of Surgery and Section of Vascular Surgery. He has completed his medical training at Stanford University, and Doctoral degree in Pharmacology at the University of North Carolina at Chapel Hill. He has served as Chief Medical Officer for a software start-up company, and has published over 25 research articles in reputed journals. His current clinical and research interests focus on “The influences of diabetes on peripheral arterial disease”. This field provides a unique appreciation of the complexity and potential gaps of knowledge in vascular pathophysiology and its end-stage complications.

Abstract:

Objectives: Over three 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 22 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: 356 patients were enrolled in the vascular surgery biobank prior to the index operation. 42 clinical variables were evaluated for each patient during the perioperative period. Vascular specimens were successfully collected for 54% (193) of patients who enrolled in the biobank. The majority of specimens collected were retrieved from the peripheral arterial system (50% carotid artery, 15% anterior or posterior tibial arteries, and 12% femoral or popliteal arteries). Each arterial specimen was sub-divided into maximally and minimally diseased portions to facilitate intra- and inter-patient biochemical and molecular analyses.

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, and subsequent experimental analysis.

Biography:

Arredondo E (BS, MSc, TPM) is a Biologist specialized in Organ, Tissues and Cell Donation from the University of Barcelona. He has participated in the implementation of liver tissue for hepatocytes isolation and SELICA clinical trials. He is currently responsible for the tissue at Research Department of DTI Foundation. His studies are based on how to develop networks to procure human tissue for research. He has been involved in several European projects related to organ donation such as the BSA project sponsored by the Council of Europe.

Abstract:

Introduction: The human hepatocytes in vitro model are the gold standard regarding a wide range of applications in biomedical research such as biological, pharmacological and toxicological studies. Researchers face many ethical and legal challenges in this area, and also sourcing high quality, fully characterized specimens able for use in bioanalytical processes.

Objectives: Aim of this study is to provide fresh human hepatic tissue fully characterized with high quality and viability criteria for hepatocytes isolation for research purposes.

Materials & Methods: The tissue was procured from patients undergoing planned liver resection surgeries due to primary or secondary tumors (living donors). The program required the involvement of health professionals in donor detection, tissue retrieval and evaluation, tissue perfusion and isolation. Hepatocytes isolation was conducted through the two-step collagenase perfusion technique and the established in-house methods. Cell viability and yield production were assessed and linked to donor clinical information. The program follows the European legislation to ensure quality, safety and traceability of all the procedures.

Results: The program linked eight procurement centers. A total of 222 donation offers were reported and evaluated; 116 donations have been isolated under a cold ischemia time of 8 hours. The 86.66% of the isolated tissues produced viability >80% and an average of 16.49 million of hepatocytes/gram were obtained.

Conclusions: Thanks to a consolidated hospital network for providing fresh tissue for research with high quality and viability criteria. As a result of the program, studies of the influence of donors’ pre-, intra- and post-operative parameters have been performed.

Biography:

Ping Zhang is currently working as Assistant Professor of Surgery in Cooper Medical School of Rowan University, USA. He has international experience includes various programs, contributions and participation in different countries for diverse fields of study.  

Abstract:

Aim: We recently described the success of a tissue engineered vascular graft (TEVG) created with autologous adipose-derived stem cells (ASC) in an animal model. The aim of this study is to investigate the effect of replacing the fetal bovine serum (FBS) with autologous human plasma (HP) within the culture medium as well as the effect of cryopreservation on graft creation and differentiation of ASC.

Methods: Human ASCs and plasma, isolated from periumbilical fat and peripheral blood, respectively, were collected from the same donors. ASCs were differentiated in endothelial growth medium supplemented with FBS (2%) vs. HP (2%). Proliferation and endothelial differentiation was measured by growth curves, MTT assay, quantitative PCR, up-take acetylated LDL, and cord formation on Matrigel.

Results: ASCs expanded in HP-supplemented medium showed similar proliferation to FBS-cultured ASCs and consistent differentiation toward an EC lineage (increases in CD31, von Willebrand factor, and CD144 message; up-take Ac-LDL and formed tube formation on Matrige). ASC were seeded into vascular scaffolds and subjected to increasing shear force within bioreactor (0-9 dynes x5d) to evaluate their use in creating a TEVG. Cryopreservation did not significantly alter ASC viability, proliferation, acquisition of endothelial characteristics, or retention after seeding onto a vascular graft.

Conclusions: This study suggests that replacement of FBS with autologous HP-a step necessary for the translation of this technology into human use does not significantly impair proliferation or endothelial differentiation of ASCs used as EC substitutes and ASCs are tolerant to cryopreservation in terms of maintaining EC characteristics and retention on a vascular graft.

Biography:

Igor I Katkov is a trained Biophysicist with more than 30 years of experience in Cryobiology and Cryogenic Engineering. In the last 5 years, 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™. He is the Chief Scientific Officer of Celltronix (San Diego, CA, USA). He has recently accepted a Professor-level position as the Head of the Laboratory at the Belgorod State University, Russia.

Abstract:

Cryopreservation (CP) and subsequent long-term storage (cryobanking) are important parts of both life science research and related industries and technologies. Slow freezing (SF) 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. Ice-free vitrification (VF) is an alternative approach to SF, which is gaining momentum for CP of oocytes, embryos, and some other type of cells. However, the traditional equilibrium vitrification (E-VF) requires high concentrations of permeable cyoprotectants that can be both chemically toxic and inflict substantial osmotic damage to the cells. As the alternative to both SF and classic E-VF methods, we have introduced the KrioBlast^™, an entirely system for hyperfast (100,000 K/min) and scalable (thousands of microliters) kinetic vitrification (K-VF) that is based on spray cooling of hermetic (closed) cryocontainers and allows to completely eliminate the Leidenfrost effect, which currently greatly impedes maximal achievable cooling rate. It is simple, robust, and can achieve VF for practically any type of cells, which makes it a permeable cryoprotectant-fee universal system as soon as the sample has the thickness below 20 mm, which comprise 90% of modern market of cryopreserved samples. Preliminary experiments have shown feasibility and very high survival (up to 90%) of cryopreserved human spermatozoa and pluripotent stem cells. Particular applications of the K-VF approach and KrioBlast^™ technology for CP and cryobanking for reproductive and regenerative medicine, husbandry, and cryopreservation of wildlife genetic resources are also discussed.

Biography:

M C Schiewe attained a BS/MS at UC Davis (1981)/LSU (1983) focused on Animal Reproductive Physiology. Working with the Smithsonian Institution/National Zoological Park and the NIH, he completed his PhD in Human Physiology in 1989 at the Uniformed Services University of the Health Sciences (Bethesda, MD). Subsequently, he performed his Post-doctoral studies at NIH/NCRR as an NSF Associate. He is currently a Scientific/Technical Lab Director for Ovagen Fertility and the California Cryobank. He considers himself to be a Comparative Reproductive Physiologist, specializing in Embryology, and has published more than 35 peer-reviewed papers and 110 scientific abstracts, including several research award presentations

Abstract:

For the past two decades, fresh and cryopreserved testicular sperm has been effectively used by sperm injection in the production of healthy IVF children. However, for many ART laboratories, the processing of testicular biopsy (TBx) tissue and isolation of viable sperm remains laborious and unreliable. Our goal was to simplify TBx handling, processing and cryopreservation procedures, while optimizing cryopreserved sperm motility by integrating pre-freeze in vitro culture (IVC) and whole tissue freezing procedures. Comparative testicular tissue IVC and evaluation was performed as part of the standard processing of whole TBx tissue (i.e., intact mass of tubules) for cryopreservation and/or for fresh use in an ICSI cycle. Two prospective studies were conducted to validate, optimize and understand the virtues of testicular tissue IVC at different temperatures (21, 30 or 37°C). Concurrently, the effectiveness of IVC-cryopreserved TBx sperm was documented with fertilization rates, clinical pregnancies and live birth data. Reliable post-thaw motility of testicular sperm was achieved by promoting pre-freeze total and progressive motility through IVC (24-96h) post-biopsy at an intermediate temperature of 30°C. Furthermore, it was determined that whole tissue cryopreservation effectively maintained post-thaw motility of IVC TBx tissue (up to 85% viability retention), with no differences in ICSI-fertilization rates or pregnancy outcomes compared to fresh TBx sperm used for women under 38 years old. Over the past 17 years, intact whole testicular biopsy cryopreservation has proven highly effective without laborious pre-freeze processing, by simply adopting IVC of TBx tissue into clinical practice. Today, the technology is also being applied to freeze preservation efforts for men undergoing cancer or vasovasotomy-related surgeries.

Biography:

Roberto Hernan, after his Bachelor´s degree in Biology (1994) worked as Research Technician at the University of Newcastle Upon Tyne (UK), focused on his cancer research career at St. Jude´s Children Research Hospital, Memphis (USA) and in 2005 obtained his PhD in Pediatric Oncology at Newcastle University. He then led the Business Development of Pharmakine during 7 years. At present, he holds the position of Chief Scientific Officer at Cellulis Ltd., where he is also a partner. He participated as a major contributor in ten scientific publications and has led the development of four different patents within the cryopreservation field.

Abstract:

Many cell therapy products need to be frozen in order to maintain product stability. Freezing and thawing cells correctly require several processes that need to be performed with care to avoid cell damage. The inadequate performance and lack of uniformity of these protocols may infer undesirable inconsistencies on the cellular products jeopardizing therapeutic efficacy. Despite current technological advances, cryopreservation protocols have been highly conserved during the last 65 years since the very first discovery of cryoprotectants. Hereby we propose a novel methodological approach to cryopreservation which introduces simple enclosed mechanisms that assure the correct standardization of freezing and thawing processes, as well as rendering a final product formulation for the cell therapy market. These mechanisms take place inside the Limbo™ vial, making the post-thawing cellular reconstitution process operator-independent. Limbo™ cryovials offer the opportunity to avoid sample washing to diminish the DMSO effect, reducing in turn expensive costs and resources at the point-of-care. Furthermore, this technology introduces a unique dry thawing system which not only enforces safe and correct thawing protocols but also eliminates contamination risks associated to water baths. In summary, this novel technology is a safeguard for most frozen cell therapies because it avoids sample handling at the point-of-care at the same time that addresses the need for appropriate cellular recovery standardization protocols in the clinic. 

Biography:

Bo He, MD, PhD now is an Associate Professor of Orthopedic and Microsurgery in the First Affiliated Hospital of Sun Yat-sen University. He started cryopreservation research since 2005, and is familiar with preservation and replantation of grafts with/without blood supply. He planned to set up biobank of various freezed tissue and organs, such as fingers, limbs, nerve and other soft tissue. He was once working in Stanford University and indulged in basic research about cryopreservation and tissue engineering. Also, he received the certification of American National Institutes of Health (NIH) for clinical research. 

Abstract:

Abstract

Background: In spite of the relatively high success rate of limb replantation, many patients cannot undergo replantation surgery because the preservation time of an amputated limb is only about six hours. In addition, although allotransplantation of composite tissues is being performed more commonly with increasingly greater success rates, the shortage of donors limits the number of patients that can be treated. So the purpose of this study is to examine the feasibility of cryopreservation and replantation of limbs in a rat model.

Methods: Sprague-Dawley rats were divided evenly into group A (above-knee amputation) and group B (Syme’s amputation). One hind limb was amputated from each rat. The limbs were irrigated with Cryoprotectant, cooled in a controlled manner to −140°C, and placed in liquid nitrogen. Thawing and replantation were performed 14 days later.

Results: In group A, the limbs became swollen after restoration of blood flow resulting in blood vessel compression and all replantation’s failed. In group B, restoration of blood flow was noted in all limbs after replantation. In one case, the rat chewed the replanted limb and replantation failed. The other rats were followed for three months with no abnormalities noted in the replanted limbs.

Conclusions: Limbs with a minimal amount of muscle tissue can be successfully cryopreserved and replanted.