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

Biography:

Christina Ellervik completed her MD in 2002 and her PhD in 2007 from University of Copenhagen, Denmark, and was board certified in Clinical Biochemistry in 2009 from National Board of Health, Denmark. From 2010-2015, she was an Associate Professor at University of Copenhagen, Denmark. Currently, she is a Visiting Scientist at Boston Children’s Hospital and Department of Preventive Medicine, Brigham and Women’s Hospital, Boston, MA, USA. She is the Founder of the Danish General Suburban Population Study (N=21,000) with 200,000 biospecimens and Co-editor for the journal, Clinical Chemistry. She has supervised several MD students for their PhD and Master’s Degree in Clinical Biochemistry.

Abstract:

Most errors in a clinical chemistry laboratory are due to preanalytical errors. Preanalytical variability of biospecimens can have significant effects on downstream analyses, and controlling such variables is therefore fundamental for the future use of biospecimens in personalized medicine for diagnostic or prognostic purposes. Currently, such preanalytical variables are not routinely documented in the biospecimen research literature. The focus of the presentation will be on preanalytical variables affecting human biospecimen integrity in biobanking, such as biological and environmental factors, collection, processing, storage, transport and retrieval. The impact of such variables on the integrity of the biospecimens as well as the interpretation of downstream analyses, costs, and FDA requirements will be discussed. It will be discussed how knowledge of preanalytical variable effects can prevent the need to waste the time and costs of repeating sample collections.

Biography:

Měřička P graduated at the Charles University in Prague, Medical School Hradec Králové in 1976. In 1979 he received specialization in Pathology and in 1989 in Tissue Banking at the Institute for Post-graduate Medical Education in Prague. In 2006, he completed his PhD in Medical Biology. He is member of the Society for Cryobiology and took part in organization of several international cryobiology meetings. He is the Head of the Tissue Bank University Hospital Hradec Králové and External Teacher at the Charles University in Prague. He is author or co-author of 80 papers dealing mostly with cryobiology and tissue banking.

Abstract:

The conditions of tissue collection and handling before start of freezing are critical for the safety and quality of transplantation of cryopreserved grafts. We analyzed risk factors, such as donor’s age (DA), time between tissue harvest and mixing with the cryoprotectant (TP), exposure to the cryoprotectant before freezing (CPAE) and the initial contamination rate (ICR) in the group of 30 arterial (AG) and 30 venous grafts (VG) collected during the multiple organ harvests in brain death donors . Only grafts meeting the criteria of release for clinical application including the proof of sterility at output control were included. The grafts were transported in the pre-cooled organ preservation solution containing gentamycin to the tissue establishment and processed in the clean rooms of the grade A with the background B. After input control and decontamination procedure each graft was put into double plastic bag filled with 50 ml of pre-cooled 10% hydroxyethlystarch solution and an equal volume of pre-cooled 20% (v/v) dimethylsulphoxide solution was added. There were no considerable differences in values (mean ± SD) of DA, TE and CPAE between AG and VG: AG/VG: DA 45±11/46±9 years, TP 20±7/19±8 hours, CPAE 31±14/30±12 minutes. The ICR was lower in VG (6.6%) than in AG (16.7%). Mycological tests were always negative. The analyzed factors do not represent danger for the graft quality. The results demonstrate advantages of using aseptic harvest procedures and document the high level of cooperation between the tissue establishment and procurement establishments.

Biography:

Mitra Mahdavi-Mazdeh has completed her MD from Tehran University of Medical Sciences and subspecialty of Nephrology from the same university. She is the Director of Iranian Tissue Bank Research Center. She has published more than 70 papers in reputed journals and has been serving as an Editorial Board Member of International Journal of Organ Transplantation Medicine and Head of Tissue and Cell Committee of Middle East Society of Transplanation.

Abstract:

Iranian Tissue Bank (ITB) has been the first multi-facility tissue bank in Iran. Its activity started in 1994 by preparing homograft heart valves and later on provision of gross bone for orthopedic surgeons. From 1994 to 2006, more than 2700 tissues from cardiac dead donors were procured. Later on, it got expertise for higher technical activities to produce particulated mineralized and demineralized bone allograft in different sizes (powder, crushed, chips and matchsticks). Different preservation techniques from simple hypothermia and freezing to lyophilization were developed which provided facilitation in transport in a country with more than 1.5 million Km². In 2012, partial split-thickness skin allograft and acellular dermis were added to the production line. At present, all kinds of tissues are provided and this center is mainly the platform of research in the field of cell seeding on human scaffolds. The future field of this center will be expanding the activity of tissue procurement in Middle East and production of semisynthetic grafts. All procedure of tissue processing is done under clean conditions of class 100 laminar flow hood in class 10000 clean room. All tissues are procured under aseptic conditions. 

Biography:

Yaffa R. Rubinstein is Program Director at the Office for Rare Diseases Research/National Center for Advancing Translational Sciences (NCATS)/National Institutes of Health (NIH), where she directs the NIH/NCATS GRDRSM Program, a Global Rare Diseases Patient Registry Data Repository, and the RD-HUB, a database for biorepositories and biospecimens, she led the development of the registry’s common data elements (CDEs) and the model informed consent for participating in patient registries. At NIH, she is an active member of the CDEs working group as well as disease-specific CDE working groups. She is also an active member of the European rare disease initiative, RD-Connect/IRDiRC, and a passionate supporter of rare disease patient advocacy groups and their families, providing them with assistance and information about patient registries and the importance of sharing data and biospecimens. She trained as a molecular biologist and received a PhD from the University of Maryland, College Park.

Abstract:

Many of the problems and difficulties associated with biospecimens for common diseases also apply to rare diseases biospecimens. In the latter, however, these problems are more acute, because of the additional challenges that uniquely pertain to research in rare diseases. Rare disease biospecimens, to the extent that they are available, are widely dispersed across geographical regions and among various government supported and private bio-repositories. 

Biorepositories (Biobanks) can provide the fuel to stimulate collaborations between patients, researchers and industry to accelerate research to develop drugs, therapeutics and, hopefully cures for these rare diseases. bio-specimens with well annotated clinical information are essential for medical research, specifically in the era of personalized and precision medicine. Because of the rarity of these biospecimens, global sharing and collaboration for standardization of high quality of samples with the associated data and interoperability between the different databases collecting patient’s samples and data is important.

Unfortunately, this effort is being hampered due to a combination of many factors which includes lack of standardization in data collection and the quality of the samples. Also lack of a consensus on human subject issues, ethical, legal regulations (informed consent, ownership, and patient privacy), interferes with global sharing of material and the associated data,

The rare disease needs, challenges and initiatives to address these hurdles will be discussed.

Biography:

Devon Kelly is a Molecular Biologist with over 16 years’ experience in specimen repository management, 10 years within pharmacogenetics research and 5 years in basic research. Currently, she focuses on standardizing oncology repository practices, including the methods by which patients are consented to donate biological samples, how the specimens are collected and stored, and in how annotative data is managed. She is a member of the International Society of Biological and Environmental Repositories, and has gained accreditation of her biobank from the College of American Pathologists.

Abstract:

There is a gap between the understanding that researchers have about clinic operations, and that of clinicians in what researchers needs are for quality tissue collection. The subsequent creation of silo workflows results in lost opportunities to gather optimal numbers of quality cases, thereby limiting research utility. With a view towards addressing this gap, the OHSU BioLibrary has been developing strategies for partnering across institutional departments, with the goal of enhancing research outcomes and fostering sustainability. The BioLibrary has developed models for interdepartmental relationships, with consideration of strategies for analyzing the impact of biobanking activities and enlarging the added value (both financial and non-financial) of these partnerships within OHSU. These models include working with clinical department chairs and principal investigators to establish Memos of Understanding that outline the responsibilities of each party in patient consenting, tissue collection, and distribution procedures (including any revenue sharing), and instating liaisons across the groups. By maintaining an interconnected presence in these disparate environments, we have been able to increase donor catchment, collection efficiency and data accuracy. Similarly, there has been an increase in the number of principal investigators that the efforts may support, and generation of value through increased publications, grants and revenues. Institutions should consider different relationship models for biobanking partners, financial sustainability and profit sharing, and strategies for analyzing the impact of biobanking activities when designing their internal processes, with a view towards bridging gaps between the clinic and research arenas.

Biography:

Daniele Mondello is the Founder of Olomedia, an Italian Company devoted to the development of Health Software Solutions. In his career, he has held various roles, as Team Leader, Project Manager and Quality Manager in the provision of engineering services. He operates in the Italian Healthcare sector providing its expertise in projects based on "Web-Based" technologies and performing an intense training activity during refresher courses in various subjects related to Information Technology (Usability, Accessibility, Semiotic, Programming Languages, Web Server, etc.). He is an assertive supporter of the importance of automation in the software development process, on which he focuses his research and free time.

Abstract:

The Sample PREanalytical Code (SPREC) is a seven-element-long code in which each element corresponds to a preanalytical variable and contains a string of letters (different for fluid or for solid tissues) in a defined order. SPREC has been released by the ISBER (International Society for Biological and Environmental Repositories) Biospecimen Science Working Group and it has been adopted in order to track and make explicit preanalytic variations in collection, preparation and storage of specimens. These features, together with software capable of organizing sample collection, which are essential for the optimal management of a biobank. Several factors are fundamental prerequisites for the development of a software platform. It has to be configurable in real time for new requirements; it has to connect professionals with each other, across hospital, to provide a multidisciplinary approach. It has to be easy to use (user friendly); it has to permit easy research in the data and generate reports; it has to give information about donors' health; and it has to give information about preanalytic variations in collection, preparation and storage of specimens. In this light, oloHEALTH_BioBim, a platform with web interface integrated with a SPREC generating tool for the management of sample requirements, has been developed. oloHEALTH_BioBim enables the traceability of the entire life-cycle of biological samples, their organization and storage; and retrieval of associated data, while ensuring full and continuous compliance with national and international regulations.

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 of the tissue for Research Department at 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:

Donation and Transplant Institute (DTI) is a non-profit foundation, founded in Barcelona (Spain) in 2008, with the goal to support the researchers by providing human biological samples with high quality standards according to project needs. DTI has created a platform that is able to coordinate the donation of human organs, tissues and cells for biomedical research purposes, which encompasses strong collaborations with national and international procurement centers. Obtaining human biological samples, from living and deceased donors, is possible thanks to the organization of a hospital network which includes donor detection, evaluation, tissue procurement and tissue processing, if needed. Along with the sample donation, demographic information and donor clinic profile are obtained to develop an extensive data base, that allows researchers to access these facilities for scientific puposes. All projects are performed according to Spanish and European legal requirements in order ensure quality, safety and traceability of the generated material. DTI is currently involved in the obtainment of liver resections for hepatocytes isolation (7468 millons hepatocytes/gram) for research purposes (2009-present) achieving a viability average of 83.5%. Furthemore, DTI is participating in the development of projects related to obtaining whole and resected livers, kidneys, condrocits, heart valves, etc. Tissues are delivered with an average cold ischemia time of 16 hours in Europe. DTI contribution makes possible to provide key data from early stage discovery through pre-clinical research, drug safety evaluation, biomarker development, patient pre-selection and entry of novel compounds into the clinic.

Biography:

Taosheng Huang is a Physician-Scientist. Currently, he is a Professor with tenure in Human Genetics, Director, Program of Mitochondrial Medicine, and Associate Director of the Molecular Diagnostic Laboratory at Cincinnati Children's Hospital Medical Center (CCHMC). He has been actively engaged in many programs in China. He is an Honorable Professor of Peking Union Medical School, a member of the special committee for Yusheng Yuyou of People’s Republic of China, Advisory Board Member to Chinese Ministry of Health for targeted therapy and a Principal Investigator for Birth Defect Control Program of Chinese Ministry of Health.

Abstract:

The genetic integrity of iPSCs is an important consideration for therapeutic application. In this study, we examine the accumulation of somatic mitochondrial genome (mtDNA) mutations in skin fibroblasts, blood and iPSCs derived from young and elderly subjects (24-72 years). We found that pooled skin and blood mtDNA contained low heteroplasmic point mutations but a panel of ten individual iPSC lines from each tissue or clonally expanded fibroblasts carried an elevated load of heteroplasmic or homoplasmic mutations, suggesting that somatic mutations randomly arise within individual cells but are not detectable in whole tissues. The frequency of mtDNA defects in iPSCs increased with age, and many mutations were non-synonymous or resided in RNA coding genes and thus can lead to respiratory defects. Our results highlight a need to monitor mtDNA mutations in iPSCs, especially those generated from older patients, and to examine the metabolic status of iPSCs destined for clinical applications.

Biography:

Lee Kerkhof is currently working as Chair in Department of Marine and Coastal Sciences, Rutgers University, USA. He has international experience includes various programs, contributions and participation in different countries for diverse fields of study. His research interests reflect wide range of publications in various national and international journals. 

Abstract:

This report describes BioDry (patent pending), a method for reliably preserving the biomolecules associated with aquatic microbial biomass samples, without the need of hazardous materials (e.g. liquid nitrogen, preservatives, etc.), freezing, or bulky storage/sampling equipment. Gel electrophoresis analysis of nucleic acid extracts from samples treated in the lab with the BioDry method indicated that molecular integrity was protected in samples stored at room temperature for up to 30 days. Analysis of 16S/18S rRNA genes for presence/absence and relative abundance of microorganisms using both 454-pyrosequencing and TRFLP profiling revealed statistically indistinguishable communities from control samples that were frozen in liquid nitrogen immediately after collection. Seawater and river water biomass samples collected with a portable BioDry “field unit", constructed from off-the-shelf materials and a battery-operated pumping system, also displayed high levels of community rRNA preservation, despite a slight decrease in nucleic acid recovery over the course of storage for 30 days. Functional mRNA and protein pools from the field samples were also effectively conserved with BioDry, as assessed by respective RT-PCR amplification and western blot of ribulose-1-5-bisphosphate carboxylase/oxygenase. Collectively, these results demonstrate that BioDry can adequately preserve a suite of biomolecules from aquatic biomass at ambient temperatures for up to a month, giving it great potential for high resolution sampling in remote locations or on autonomous platforms where space and power are limited.

Biography:

Michael L Moeller, PhD is the current Chief Scientist of American CryoStem Corporation, where he has applied his nearly 20 years of work in Human Tissue-derived Stem Cell Biology to develop and commercialize new reagents and technical platforms for the collection, expansion, differentiation, banking and application of human adipose-derived mesenchymal stem cells. 

Abstract:

Autologous stem cell therapies permit a therapeutic approach devoid of the possibility of transplant rejection, the need for immunosuppressive therapies or any potential problems which might arise from transmitted pathologies. It is also attractive as it offers a “self-healing-self” paradigm that is appealing to many. It has been long been known that mesenchymal stem cells (MSCs) are largely resident in the perivascular niche surrounding blood vessels throughout the human body. Any tissue that is highly vascularized can conceivably serve as a robust and reliable source of MSCs, including adipose tissue obtained from liposuction procedures. In this presentation, we will look at American CryoStem, a small but growing biotech focused on the collection, expansion, and banking of client-specific adipose tissue and mesenchymal stem cells. 

Biography:

Xiaoling Chen has completed her MA from Graduate School of Chinese Academy of Agricultural Sciences. She is Professor of Institute of Crop Sciene, Chinese Academy of Agricultural Sciences and is in charge of crop germplasm resources in vitro conservation and cryopreservation research work in China National Crop Genbank. She has published more than 26 papers in reputed journals and has got 5 patents. She has been a member of the Society for Cryobiology.

Abstract:

A national crop germplasm resources conservation system, including in situ conservation and ex situ conservation, has been preliminarily built up in China. Up to now, 169 in situ conservation sites have been established in provinces, municipalities and autonomous regions. All these in situ conservation sites involve wild relatives of crops, such as wild rice, wild soybean, wild wheat relatives and so on. Ex situ conservation system includes one national long-term genebank, one national duplication genebank, 10 national medium-term genebanks, 43 national field repositories and two in vitro genebanks for potato and sweet potato, respectively. Around 60,649 accessions are in field repositories and 14 species, 2,091 accessions of potato cultures and 16 species, 1,198 accessions of sweet potato cultures are in in vitro genebanks. The National Crop Genebank of China is the long-term and mid-term preservation center and advanced research center on conservation technologies for crop genetic resources in China. It includes one long-term genebank and one medium-term genebank. It holds crop seeds of 898 species, 404,690 accessions for long-term at -18°C and 212,692 accessions for mid-term at -4°C, respectively. There are 38 species, 353 accessions of cultures in in vitro at 10-20°C and 15 species, 140 accessions of pollen, shoot tips and dormant buds or segments cryopreserved at -196°C in liquid nitrogen for vegetatively propagated crops.

Biography:

Kruthika Vinod T P completed her PhD Neurochemistry in 2008 from NIMHANS, Bengaluru, India, a premier institute for Neurosciences in India. In 2009, she joined as Senior Scientific Officer in Department of Neurochemistry and was given the responsibility of screening metabolic disorders. Her research interest includes cerebral venous thrombosis, metabolic disorders and prenatal screening of metabolic disorders. She has 12 international publications in peer-reviewed journals. She has received young physicians/scientists awards in Neurology conferences and many travel fellowships. Recently, her student received “Chen’s student award” for outstanding research for her work on GA-I at 11^th APCHG, Hanoi, Vietnam 2015.            

Abstract:

Dried blood spots (DBS) are important form of bio-sampling, where DNA can be stored and used for genetic studies. This has necessitated to develop an efficient and economical genomic DNA (gDNA) extraction protocol from these sampling cards. We have developed a novel and non-hazardous method called “single lysis-salting out (SLSO) protocol to extract gDNA from DBS samples. The efficiency of this protocol was evaluated against the commercial kits (Qiagen and Analytica Jena). For the purpose of this study, whole blood DBS samples were collected from 10 clinically healthy volunteers and 30 confirmed Glutaric Aciduria Type I (GA-I) patients from India. The gDNA was extracted from the collected DBS samples by SLSO, QIAamp® gDNA Micro kit and innuPREP forensic kit methods. The yield and quality of gDNA obtained was determined by measuring the absorbance using Nanodrop spectrophotometer. It was observed that SLSO method showed four-fold and eight-fold increased yield of gDNA in healthy volunteers and patient samples respectively compared to commercial kits (p<0.0001). The purity of gDNA was concordant with the commercial kits(r² ≥ 0.9). This method was found to be cost effective, reducing the per sample cost to nearly half. The suitability of SLSO method for genetic studies was confirmed by performing R402W genotyping by RFLP in GA-I patients from India. The genotyping results showed the presence of R402W mutation in 20% (6/30) of patients. In conclusion, the SLSO protocol was found to be dependable, inexpensive, non-hazardous and appropriate for genetic studies.

Biography:

Kruthika Vinod T P completed her PhD Neurochemistry in 2008 from NIMHANS, Bengaluru, India, a premier institute for Neurosciences in India. In 2009, she joined as Senior Scientific Officer in Department of Neurochemistry and was given the responsibility of screening metabolic disorders. Her research interest includes cerebral venous thrombosis, metabolic disorders and prenatal screening of metabolic disorders. She has 12 international publications in peer-reviewed journals. She has received young physicians/scientists awards in Neurology conferences and many travel fellowships. Recently, her student received “Chen’s student award” for outstanding research for her work on GA-I at 11th APCHG, Hanoi, Vietnam 2015.

Abstract:

Dried blood spots (DBS) are important form of bio-sampling, where DNA can be stored and used for genetic studies. This has necessitated to develop an efficient and economical genomic DNA (gDNA) extraction protocol from these sampling cards. We have developed a novel and non-hazardous method called “single lysis-salting out (SLSO) protocol to extract gDNA from DBS samples. The efficiency of this protocol was evaluated against the commercial kits (Qiagen and Analytica Jena). For the purpose of this study, whole blood DBS samples were collected from 10 clinically healthy volunteers and 30 confirmed Glutaric Aciduria Type I (GA-I) patients from India. The gDNA was extracted from the collected DBS samples by SLSO, QIAamp® gDNA Micro kit and innuPREP forensic kit methods. The yield and quality of gDNA obtained was determined by measuring the absorbance using Nanodrop spectrophotometer. It was observed that SLSO method showed four-fold and eight-fold increased yield of gDNA in healthy volunteers and patient samples respectively compared to commercial kits (p<0.0001). The purity of gDNA was concordant with the commercial kits(r2 ≥ 0.9). This method was found to be cost effective, reducing the per sample cost to nearly half. The suitability of SLSO method for genetic studies was confirmed by performing R402W genotyping by RFLP in GA-I patients from India. The genotyping results showed the presence of R402W mutation in 20% (6/30) of patients. In conclusion, the SLSO protocol was found to be dependable, inexpensive, non-hazardous and appropriate for genetic studies

Biography:

Xiaoling Chen has completed her MA from Graduate School of Chinese Academy of Agricultural Sciences. She is Professor of Institute of Crop Sciene, Chinese Academy of Agricultural Sciences and is in charge of crop germplasm resources in vitro conservation and cryopreservation research work in China National Crop Genbank. She has published more than 26 papers in reputed journals and has got 5 patents. She has been a member of the Society for Cryobiology

Abstract:

A national crop germplasm resources conservation system, including in situ conservation and ex situ conservation, has been preliminarily built up in China. Up to now, 169 in situ conservation sites have been established in provinces, municipalities and autonomous regions. All these in situ conservation sites involve wild relatives of crops, such as wild rice, wild soybean, wild wheat relatives and so on. Ex situ conservation system includes one national long-term genebank, one national duplication genebank, 10 national medium-term genebanks, 43 national field repositories and two in vitro genebanks for potato and sweet potato, respectively. Around 60,649 accessions are in field repositories and 14 species, 2,091 accessions of potato cultures and 16 species, 1,198 accessions of sweet potato cultures are in in vitro genebanks. The National Crop Genebank of China is the long-term and mid-term preservation center and advanced research center on conservation technologies for crop genetic resources in China. It includes one long-term genebank and one medium-term genebank. It holds crop seeds of 898 species, 404,690 accessions for long-term at -180C and 212,692 accessions for mid-term at -40C, respectively. There are 38 species, 353 accessions of cultures in in vitro at 10-200C and 15 species, 140 accessions of pollen, shoot tips and dormant buds or segments cryopreserved at -1960C in liquid nitrogen for vegetatively propagated crops.

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:

Miyamoto Keiko is a student in the Medical Communication Department at Kyoto University Graduate School of Medicine, Japan. She is interested in the public understanding of science, risk communication, and social relations.

Abstract:

This study examined residents’ attitudes toward an ongoing, real genome cohort study based on a community. The Kyoto University Graduate School of Medicine and the city of Nagahama initiated the Nagahama study for comprehensive human bioscience to promote the health of the citizens of Nagahama, develop community-based genomic-epidemiologic studies, and create a biobank. The Nagahama city government enacted an ordinance to manage the study. The mission was to prioritize the dignity of citizens. After the launch of the genome cohort study, in November and December 2009, a self-administered questionnaire survey was conducted with 2500 randomly sampled residents aged 30-74 years living in Nagahama, Japan. Responses were received from 1363 people, 187 of whom had already participated in the study. Although the local government and researchers disseminated information through leaflets and citizen-information papers to every household, sent notices by personalized letter, and held symposia and other meetings, 66% of males and 47% of females first became aware of the study when they received our questionnaire. Half of the participants understood it as a medical study involved genetic analysis. Their attitudes were significantly associated with the desire for an extensive health check-up. Particularly, positive aspects were associated with a high health consciousness in males and participation in community activities in females. Although promoting a community-based genome cohort study requires a huge effort, popularizing it is essential. Actions are vital for both monitoring public awareness and attitudes at a community level and for keeping the channels of communication open. 

Biography:

Lili E Ehrlich is a PhD student in the Department of Mechanical Engineering at Carnegie Mellon University, and is advised by Professor Jonathan A Malen and Professor Yoed Rabin. She has completed her BS from The Cooper Union, New York, NY. and MS from Carnegie Mellon University, Pittsburgh, PA.

Abstract:

Thermal conductivity of the cryoprotective agents (CPAs) cocktail DP6 (3M DMSO and 3M propylene glycol) is measured in this study, in combination with a battery of synthetic ice modulators (SIMs). The DP6 is investigated as a vitrification promoting cocktail, while the SIMs are added to suppress ice growth and, thereby, to widen the thermal conditions applicable to cryopreservation by vitrification. Key SIMs under investigation are 6% 1,3 Cyclohexanediol, 6% 2,3 Butanediol, and 12% PEG400. In addition, the thermal effects of EuroCollins as a vehicle solution on the CPA-SIM cocktail are also investigated. Thermal properties data for CPAs and SIMs are key ingredients for simulations of cryopreservation by vitrification of large-size specimens. Such simulations can be used for thermal analyses of experimental results, as well as for the design of new solutions and protocols which favor glass formation. A typical thermal protocol for thermal conductivity measurements in the current study combines a cooling rate in the range of -2.5°C/min to -50°C/min, a storage temperature at either -130°C or -180°C, and rewarming in an average rate of 3°C/min above glass transition. Thermal conductivity measurements are taken during rewarming, using a hot wire technique. Vitrification was verified by means of a scanning cryomacroscope-a proprietary device used for visualization of large-scale cryopreservation. In general, the thermal conductivity during vitrification monotonically and moderately decreases with the decreasing temperature, while during crystallization the thermal conductivity increases with decreasing temperature.

Biography:

Prem K Solanki is a Master’s degree student at Carnegie Mellon University and is advised by Professor Yoed Rabin. He has completed his Bachelor in Mechanical Engineering from the Birla Institute of Technology and Science in India. His primary interest includes Heat Transfer and he has previously worked on projects entitled “Applications in advanced manufacturing systems, passive cooling systems, and specialized vehicle systems”.

Abstract:

Aim: This study focuses on the analysis of thermo-mechanical stresses in large-size cryobags, with the long-term goal of predicting the likelihood of fracture in vitrification processes. Previous studies have demonstrated that the maximum level of stress is reached at the early stages of rewarming, which outlines the current path of investigation.

Method: The current study focuses on a pillow-shaped cryobag, having typical dimension of 61 mm×61 mm and a variable thickness of up to 61 mm, representing variable content volume-cryoprotective agent (CPA). This study uses the finite elements analysis software ABAQUS to simulate the coupled thermo-mechanical problem. This study uses the CPA cocktail VS55 as a representative cryopreservation medium, which has been well characterized by our research team in recent years.

Results: For a single stage rewarming, the maximum stress was found at the volumetric center of the cryobag. However, other thermal protocols may lead to a maximum stress at the edge of the bag. The maximum stress decreased between 5.5% and 79%, depending on geometric parameters, when an intermediate temperature-hold period at -110°C was introduced during rewarming (about 10°C above glass transition). It is further demonstrated that the mechanical stress can be dramatically reduced with the incorporation of volumetric heating, such as nanoparticles in an oscillating magnetic field.

Conclusions: From a thermo-mechanical perspective, it would be advisable to fill the cryobag with the minimum amount of CPA and to maintain as flat as possible. Volumetric heating can reduce rewarming-phase thermo-mechanical stress.

Biography:

Zyiad Mushref Abu Khaizaran is a PhD student in the Department of Biology and Biotechnology from Bethlehem University and Palestine Polytechnic University, Palestine. He has completed his Master of Biotechnology from Palestine Polytechnic University - Palestine and BSc of General Science from Al-Quds Open University, Ramallah, Palestine.

Abstract:

Modern dairy cattle breeding strategies depend on linkage analysis and quantitative trait loci (QTL) of genes involved in milk yield and composition. This is because of their biological desired quantitative traits that play key roles in milk production. In this study, three genes directly related to milk production: Prolactin (PRL), bovine kappa-casein (K-CN) and the pituitary-specific transcription factor (PIT-1) were analyzed in 144 cows. The aim of this study was to identify polymorphisms in the Holstein-Friesian cattle breed in Palestine in relation to the genetic markers and allelic variants of the three genes. Collection of samples depended on an experimental design that was completely randomized (CRD) and blood samples were collected from different cities across the West Bank, Palestine. The genotypes were determined through the Polymerase Chain Reaction-Restriction Fragments Length Polymorphism (PCR-RFLP) technique. The amplified fragments of PRL (294-bp), K-CN (530-bp) and PIT-1 (451-bp) were digested with RsaI, HindIII and HinfI, respectively. Statistical analysis found that the prolactin allelic substitution (AG, GG) played a role in milk production with a p-value of 0.00643 and α (0.001**), the AG allele of PRL being more favorable for milk production as compared to the GG allele. Genetic variants of the bovine K-CN gene played a role in milk production with a p-value of 0.04071 and α (0.01*), the AA allele possessing more positive effect than the BB and AB alleles. Similarly, the allelic substitution of the PIT-1 gene affected milk production with a p-value of 2.274e-05 and α (0***), the AA allele exercising a more positive effect followed by the AB and BB alleles, respectively. Among the three studied breeds (Friesian, hybrid and local), results show that the Friesian breed possesses higher overall milk production in Palestine as compared to the other two breeds.

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.