Day 2 :
Biography:
I worked at the Roslin Institute for 34 years and was involved in the Cloning project. I was the embryologist who cloned the first animals from cultured cells Morag and Megan which was the enabling technology which led to the birth of Dolly the sheep the following year. The project then progressed to produce cloned sheep which produced “human blood factor 9”. Subsequently I used my skills as an embryologist to develop several methods of producing transgenic animals. I took early retirement from Roslin and travelled extensively teaching some of the techniques I had developed at Roslin. Notable projects have been to act as consultant on the project in Dubai to produce the first cloned camel. I also acted as consultant to a project in Kenya producing the first cloned native breed of cattle.
Abstract:
Mammalian cells as well as gametes and embryos have been stored frozen under liquid Nitrogen for many years. Sperm was first cryopreserved around 1953. Freezing and storage of tissues and cells in liquid nitrogen at -196â°C prevents biological activity of the tissue including activity which would result in cell death. Storage under liquid nitrogen is thought to provide indefinite safe storage so long as the temperature does not rise above the “Glass Transition” point which occurs at -136â°C, this is the temperature where biological activity is seen to start occurring and so deterioration begins.
Can tissues be stored safely in a dry state without damage occurring to the cells? If this can be perfected it would cut the cost of biobanking considerably!
Keynote Forum
Youhe Gao
Beijing Normal University, China
Keynote: Saving Urinary Proteins and Nucleic Acids on a Piece of Membrane Simply and Economically
Time : 10:35-11:05
Biography:
Youhe Gao is a Professor in Beijing Normal University. He received his MD from Peking Union Medical College and his from PhD from University of Connecticut and Postdoctoral training from Beth Israel Deaconess Medical Center, Harvard Medical School. He was the Professor of Department of Pathophysiology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences/ Peking Union Medical College from 2001-2014. His research interests include biomarker discovery in urine, urine biobanking, protein interaction methods and related bioinformatics.
Abstract:
Urine has become more and more recognized as important biomarker source in biomarker discovery. By nature, urine accumulates all kinds of changes and changes are the most basic property of biomarker. In this sense, theoretically urine can be even better biomarker source than blood. There may be a urine biomarker era ahead of us. Even though metabolites were analyzed extensively as potential biomarker in urine, proteins and microRNAs may possess more specificity of the diseases. Saving proteins and nucleic acids were challenging because they not only took huge space but also gradually degraded even frozen in -80 degree. Here we present a simple and economical method that make storing large number of urinary proteins and nucleic acids clinical samples possible. Urine samples were filtered through the membrane and urinary proteins were adsorbed onto the membrane. The membrane used can have high affinity to proteins or nucleic acids. Then the membrane was dried and stored in a vacuum bag which kept the protein and microRNA faithfully preserved. The membrane may even permit storage at room temperature for weeks. This simple and inexpensive method requires minimal sample handling, uses no organic solvents, and is environmentally friendly. With this method, large number of samples will be available to biomarker discovery and validation. Biomarker research can be significantly more efficient. There has not been any new form of tissue preservation for years. This membrane, which we call urimem, may be kept with medical record of all consenting people in the future
- Track 1: Human cancer biobank
Track 4: Cryopreservation
Track 8: Biobanking in microbiology
Track 10: Neuro Biobank
Session Introduction
Domenico Amato
AL.CHI.MI.A. S.r.l, Italy
Title: AL.CHI.MI.A. S.r.l. – Human tissue processing specialists.
Time : 11:20-12:00
Biography:
Domenico Amato is a sales manager in the AL.CHI.MI.A. S.r.l. company. Since 2011 the main task of Domenico Amato was the management of the markets in EMEA territories. He graduated in biology science at “La Sapienza” university of Rome in 2006. Also provides technical & scientific support to the sales.
Abstract:
AL.CHI.MI.A. S.r.l. is made up of experts from pharmaceutical and medical device industries. It was established in 1993 and it is based in Padova, Italy. Since then AL.CHI.MI.A. S.r.l. is dedicating all research efforts to medical devices. Currently, we carry three product lines: Eye Banking, Ophthalmic Surgery and Tissue Banking. However, in recent years our expertise has grown especially for the field of human tissue banks. The result has been a thriving scientific production; on average a new paper per year and participation to major European congresses related to human tissue banking. All this is then materialized in the design of new medical devices exclusive for processing human tissue as decontamination (BASE 128) and microbiological analysis (RESEP Syringe, ready for sale). At cooperates with several banks of human tissue in Europe, all this for garatire to products especially for high standards required necessities; providing products CE marked and designed for use with human tissue.
Koichi Kyono
Kyono ART Clinic, Japan
Title: Safety of frozen-thawed ovarian tissue (FTOT) auto-transplantation following cryopreservation in terms of minimal residual diseases for cancer patients and its accuracy of analysis
Time : 12:00-12:25
Biography:
Koichi Kyono received his MD from Fukushima Medical College in 1978 and his PhD in obstetrics and gynecology (reproductive biology) from Tohoku University in 1984. He was a member of the Tohoku University team which achieved the first IVF pregnancy and delivery in Japan in 1983. He is currently the President of Kyono ART Clinic in Sendai and Tokyo. His long-term research interests include studies on cryopreservation (oocytes, ovarian tissue and testicular spermatozoa) in vitro culture, andrology (Micro-TESE, IMSI) and endocrinology (ovarian stimulation).
Abstract:
Approximately 30 babies have been born following auto-transplantation of FTOT from cancer patients. We reported ovarian metastasis (OM) by histology of 5,571 autopsy findings of females under the age of 40 in Japan. The percentage of OM was the highest in those with gastric carcinoma (55.8%) followed by colon cancer (26.6%) breast cancer (24.2%) pulmonary carcinoma (23.4%) malignant lymphoma (13.3%) uterine carcinoma (13.1%) and leukemia (8.4%). Bittinger et al (2011) demonstrated ovarian involvement in stage III Hodgkin lymphoma. The Belgian team detected malignant cells by histologic evaluation in 2 (6%) out of 32 patients with non-Hodgkin’s lymphoma, in the medulla (1/32) and in the cortex (1/32). Furthermore, the incidence of OM in breast cancer patients is common in advanced-stage. Dolman et al. (2012) evaluated the presence of residual leukemic cells in cryopreserved ovarian tissue by histology and quantitative reverse-transcribed polymerase chain reaction (RT-PCR). Malignant cells were found in 2 of 6 CML patients and 7 of 10 ALL patients by RE-PCR though, these were not found by conventional histological evaluation. Abir et al. (2010) evaluated ovaries from 8 patients with Ewing sarcoma by histology,　immunohistochemistry and RT-PCR. No ovarian involvement was found in histology but RT-PCR analysis revealed the presence of cancer cells in one patient. Therefore, FTOT auto-transplantation following cryopreservation should not be performed if malignant cells are found by histology or RE-PCR. We recommend both histology and RT-PCR before FTOT transplantation in cancer patients.
Clelia R A Bertoncini
University of Sao Paulo, Brazil
Title: Regeneration of brain tissue by mesenchymal stem cells in a stroke model involves both raised nitric oxide and VEGF as decreased oxidative stress and apoptosis to healthful levels
Time : 12:25-12:50
Biography:
Clélia Rejane Antonio graduate at Quimica Licenciatura from Universidade Federal de Santa Catarina (1986), master's at Físico-Quimica from Universidade Federal de Santa Catarina (1989) and ph.d. at Bioquimica from Universidade de São Paulo (1995). Has experience in Biochemistry, focusing on Biochemistry, acting on the following subjects: transgenicos, iron, dna, oxidative stress and dna damage.
Abstract:
Mesenchymal stem cells (MSCs) hold tremendous potential for tissue regeneration. We hypothesized that the action of these cells is not only to repopulate the area of brain damage, but mainly to secrete neurotrophic and proliferative factors which could induce or stimulate the recovery of the damage. As we previously described, the model Stroke Prone Spontaneously Hypertensive Rat (SHRSP) exhibits a hippocampal damage, which can fully be recovered after transplantation of MSCs. Moreover, such cell repopulation occurred whereas apoptosis, superoxide and lipid peroxidation were reduced to normal levels, as verified in parallel with helpful Wistar Kyoto (WKY) controls (Calió et al; Free Rad Biol Med, 2014). The objective of this work is to investigate if transplanted MSCs could be involved in proliferation of neural cells by analysis of the levels of nitric oxide (NO) and expression of vascular endothelial growth factor (VEGF).
Methods We performed a comparison of the brains isolated from SHRSP treated or not with MSCs with those from normotensive Wistar Kyoto (WKY) controls by using quantitative RT-PCR, immunohistochemistry and biochemistry assays. MSCs were obtained from the femur and tibiae of 12-week-old WKY rats and characterized for the presence of mesenchymal surface antigens (CD90 and CD105) and the absence of hematopoietic markers (CD45, c-kit, and Sca1) by flow cytometry before the transplantation. 1x106 MSC previously labeled with CFSE were injected into cistern magna through the atlanto-occipital membrane of 48-week-old SHRSP. The expression of VEGF was assayed both by Western blot and real-time (quantitative) RT-PCR analysis. β-Actin was used as a housekeeping gene. Nitric oxide (NO) was measured to evaluate its possible role in neural proliferation or endogenous brain cells regeneration after MSCs transplantation to the brain of stroke rat models. The quantification of generated NO was based on the gas-phase chemiluminescent reaction between NO and ozone; data were detected by the Nitric Oxide Analyzer (Sievers Instruments, Boulder, USA), a high-sensitive detector (~1 pmol) of NO in liquid samples.
Results An increase of almost threefold of VEGF expression was observed in the MSC-treated SHRSP group, thereby suggesting that transplanted stem cells have a proliferative potential by inducing proliferation of neural stem cells. Similar results were obtained in terms of elevation of generated NO detected by chemiluminescence. Thus, we suggest that both Vegf and NO can contribute to the recovery of hippocampal damage associated with oxidative stress and apoptosis in the spontaneously hypertensive stroke model SHRSP.
Conclusion: Our data suggests that MSCs could secrete or induce the same types of proliferation factors, such as Vegf and NO, which could stimulate the recovery of brain damage by endogenous stem cells. Thus, stem cell transplantation is a promising therapy, which could provide trophic support, helping promote survival, migration, and differentiation of endogenous precursor cells.
Supported by Fapesp, CNPq and FAP-Unifesp
Aline Azevedo
Universidade Federal de São Paulo, Brazil
Title: Human embryo cryopreservation: Missing the fear
Time : 13:35-14:00
Biography:
Biomedic from the University Center Uni FMU (2003). Master degree-Medicine Department of Nephrology-Federal University of São Paulo-UNIFESP (2006). PhD by Ginecology Department-UNIFESP and Cleveland Clinic ( 2011). Pos -doc by Department of Gynecology-UNIFESP ( in progress). Young Scientist Award in 2011 (14th World Congress on Controversies in Obstetrics, Gynecology and Infertility-Paris, France). Now, acting as embryologist and Researcher at the Federal University of São Paulo. She has experience in Biochemistry, with emphasis in Human Reproduction, mainly following themes: oxidative stress, antioxidants, superoxide, Endometriosis.
Abstract:
Background Embryo cryopreservation is now a routine procedure in assisted reproductive laboratories. Currently, is more important than ever for the cumulative pregnancy rate after In vitro Fertilization (IVF) . Recently, increases in success rates after frozenthawed embryo transfer (FET) are nearing the success rates of fresh embryo transfer (ET) and this can encouraging the use because reduces risks like low birth weight and prematurity, ovarian hyperstimulation syndrome (OHSS), among others. Furthermore, the controlled ovarian hyperstimulation affects the endometrial maturation.
Objective To evaluate ongoing pregnancy rates after FET cycles and compare with the success of ET described at the literature.
Materials and Methods A retrospective study of 72 patients under IVF treatment that where indicated cryopreserved all embryos because of OHSS risks. The patients parameters evaluated was age, infertility factor, number of retrieved and mature oocytes, fertilization rate, embryos transferred per patient, pregnancy, implantation and abortion rate.
Results The average age was 30.5 years; male factor was indicated in 42% of cases. The number of aspirated oocytes per patient was 28,11 which 74,35% were mature. The fertilization rate was 80,53%, the average of embryos transferred was 2.78 per transfer, the average number of transfer per patient was 1,8. The pregnancy rate per transfer was 49.59% and per patient was 83.3%. The implantation rate was 35.7%. Abortion rate was 13.89%.
Conclusions Until now, there is no consensus between different groups around the world about the best protocol, day of embryo cryopreservation, freezing method, selection criteria for which embryos to freeze, method of embryo thawing and endometrial preparation for transfer of frozen-thawed embryos. However, it has been reported a greater implantation and pregnancy rates with FET when compared with ET, suggesting superior endometrial receptivity in the absence of ovarian stimulation. Ours results agrees with recently reported data and emphasizing that we can use frozen embryos without fear. High-quality randomized controlled trials should be pursued to find out which cryopreservation protocol is the best and when will be the time to completely abandon fresh embryos transfer.
Pan Jing
Biobanking Center for Chinese Preterm Clinical Research Consortium, China
Title: Genomic and epigenomic studies of spontaneous preterm birth with an pre-banked existing Chinese cohort
Time : 14:00-14:25
Biography:
Jing Pan joined the faculty in 2013 as a Senior Lecturer. Her current instruction duties include teaching Eukaryotic Molecular and Cell Biology (BIOL3102) and RNA world (BIOL6V29). Dr Pan received a B.S. (1998) from Nankai University in Tianjin, China, and a Ph.D. (2004) in Biochemistry, Molecular, and Cell Biology from Cornell University in Ithaca, NY. Her PhD work focused on the study of spindle checkpoint, a cell cycle surveillance mechanism, in budding yeast. After her PhD, she completed postdoc training at Memorial Sloan-Kettering Cancer Center in New York City (2004-2008), where she worked on meiotic recombination in budding yeast and in mice; and at University of Texas, Southwestern Medical Center in Dallas (2009-2013), where she worked on miRNA modulation of cell polarity.
Abstract:
Spontaneous preterm birth (sPTB), consisting of preterm premature rupture of membrane (PPROM) and spontaneous preterm labor (sPTL), is the leading cause of neonatal matality and life time mobility. Genetic and genomic variations, environmental factors, and gene-environment interaction(s) are the common etiological factors involved in many physiopathological pathways of PPROM. Many studies have identified the presence of single nucleotide variations (SNVs), copy number variations (CNVs), and DNA methylations as the evidence that genetic/genomic factors and gene-environment interactions play predisposition roles in PPROM. Studies also showed that the human genome transcribed thousands of regulatory non–protein-coding RNAs (ncRNAs), including microRNAs (miRNAs) and long ncRNAs (lncRNAs), are expressed during pregnancy in placentas in addition to fetal amniochorionic membranes, and are involved in sPTB. Previously, we have reported that lncRNAs and mRNAs were differentially expressed in human embryo sacs that contain chorionic villi and maternal decidua in spontaneous abortion within the first trimester, indicating that lncRNAs have played a regulatory function in the early stage of pregnancy. We hypothesized that prenatal exposure to infection and inflammation could cause epigenetic modifications in the pathogenic pathway(s) of sPTB. We therefore undertook a study to identify genome-wide differential expression profile (DEP) of lncRNAs and mRNAs and of DEP of miRNAs and methylation, in addition to exome sequencing, with an existing banked sPTB cohort. Our studies have identified numerous differentially expressed lncRNAs and mRNAs associated with sPTB. Several pathogenic pathways constructed from the differentially expressed lncRNAs and mRNAs showed a significant association with sPTB. We further characterized lncRNAs may regulate gene transcription of collagen-ubiquitin-proteasome (CUP) in human placentas derived from PPROM. Our studies provided evidence that epigenetic modification are involved in pathogenesis of adverse pregnancies, which help in obtaining a greater understanding of the pathogenic pathways associated with sPTB, the novel information about the epigenetic mechanisms underlying sPTB, and help improve the outcome of pregnancies.
Osman Zin AlAbdin
King Saud University Medical City, Saudi Arabia
Title: A biological bank of liver samples and related information intended for hepatocellular carcinoma research
Time : 14:25-14:50
Biography:
Osman Zin AlAbdin has completed his Msc. in Biotechnology at the age of 22 years from McGill University. He has worked as a Research Assistant for 6 years at the Research Institute of Cancerology and Immunology, University of Montreal. He is presently the biobank manager at the Liver Disease Research Center. He has been serving as a member of the Tissue Allocation committee at KKUH since 2013.
Abstract:
Hepatocellular carcinoma (HCC) is among the top 5 cancers in Saudi Arabia. We evaluated the feasibility of establishing a longitudinal cohort of the patients with various liver diseases at increased risk for HCC, namely HCV, HBV, NAFLD, and liver adenoma. We have outlined the ethical, methodological and technical issues of the biobank establishment process. As a result, we have aligned with the Biobank Resource Center (developed by the Canadian Tissue Repository Network in partnership with the UBC office of Biobank education and research) in order to adapt standardized SOPs and eliminate the numerous variables that can emerge around the process of collecting surgical tissues for research. The aim of our biobank is to provide research groups with a platform of web-based socio-demographic information, detailed longitudinal clinical data couples with high quality biological samples to properly interpret research data and ultimately promote the advancement of liver disease research. Singe database (SOLID) commenced in early 2010 and collected a total of 949 HCV, 1904 HBV, 403 NAFLD and 7 Primary liver neoplastic. In addition to banking fresh frozen liver tissue, the bank contains blood in which it is processed into serum, plasma, buffy coat, RNA and DNA extraction along with subcutaneous fat, visceral fat and abdominal muscle specimens from NAFLD patients. The methods, in which the fresh liver tissue is harvested for biobanking and in which the liver is sampled for histological assessment, correlates with the strict policies, procedures and appropriate controls that are adapted according to the Biobank Resource Center best practices. A well-developed biobank is a critical prerequisite for high-quality research. This review provides an outline of certain critical elements that would need careful attention as a liver disease biobank is developed.
Beena Kumar
Monash Health, Australia
Title: Role of immunohistochemistry and biobanking in targeted cancer therapy
Time : 14:50-15:15
Biography:
Role of immunohistochemistry and biobanking in targeted cancer therapy Dr. Beena Kumar, Deputy Director, Anatomical Pathology, Monash Health, Victoria, Australia
Abstract:
Chemotherapy which has been the standard therapeutic regimen for cancer has the disadvantages of conforming to the “one size fits all” style. These standard drugs fail to distinguish malignant versus normal tissue, thus bringing along a range of adverse effects. Targeted treatment on the contrary show a greater selectivity for tumor cells and causes less damage to normal cells. It is to be noted that morphologically distinct tumours show variable biological characteristics and response to treatment. It is thus becoming important to identify these targets within the cancer tissue which include the tumour cells and the tumour microenvironment (ie, stromal cells, microvessels, and host’s immune cells), all of which could serve as potential treatment targets. Most of these biomarkers were earlier detected by molecular techniques. These tests are expensive and not easily accessible. Immunohistochemistry is an excellent surrogate to identify the proteins / targets in question. Though widely used, this technique comes with its share of challenges which could be at the preanalytical, analytical and post analytical levels. It is highly mandatory to establish robust methodologies within the laboratory to obtain the right answer, which would ultimately benefit patient management / response to treatment. The development of new treatments or diagnostics is facilitated through biomedical research which has the potential to significantly improve patient outcomes. With the advent of personalised medicine and genomic medicine, the different arms of research such as basic research, translational research, clinical research and clinical practice have merged .
Elke Smits
Antwerp University Hospital, Belgium
Title: Tumour biobank for advancing translational research in oncology
Time : 15:15-15:40
Biography:
Elke Smits earned her Bachelor of Science in Chemistry from the Catholic University of Leuven, a Master of Science in Biotechnology and a PhD in Veterinary Sciences in 1998 from the University of Gent. Elke Smits joined Devgen Inc, a spin-off company in Gent, as manager molecular cell biology for target discovery and drug development projects. In 2004, she became senior scientist at the Flemish Science Policy Council, the advisory body for the Flemish government concerning science and innovation policy. She has published over 20 peer-reviewed articles, holds several patents, wrote many policy advices and recommendations and authored the study serie Technology and Innovation in Flanders: Priorities. Prof. dr. Elke Smits currently heads the Science & Innovation department of the Antwerp University Hospital and has gained extensive experience in merging translational and clinical research within clinical practice. She holds a visiting professorship position at the Faculty of Medicine of the University of Antwerp and is liaison officer CRC Antwerp for the Center for Medical Innovation.
Abstract:
Prevention, treatment and care of cancer patients largely depend upon improvements in scientific research. Development and enhancement of cancer research networks was recommended in the Belgian National Cancer Plan, which was launched in 2008 by the Belgian Federal Ministry of Health. The objective is to create a virtual inter-institutional tumour biobank to promote translational research and to create a network for future academic, medical and/or industrial collaborations. The tumour biobank initiative at the Antwerp University Hospital (Universitair Ziekenhuis Antwerpen, UZA) (tumour biobank@UZA) will attempt to integrate into the multidisciplinary cancer care with linkage to medical and clinical data. The tumour biobank at the Department of Pathology, UZA, works according to standard procedures for sample processing and storage of fresh and paraffin-embedded tissue samples. The good collaboration and communication between the operating-room and the pathology department, and the use of international acknowledged guidelines enable the preservation of high-qualitative samples. Following preservation, 1% of the collected samples will be annually subjected to a quality control, which focuses on several parameters: sample identity, location, diagnosis, sample data, RNA integrity and concentration. Consent concerning the use of residual tissue for scientific purposes is obtained prior to surgery in accordance with the rules of the local ethics committee. The tumour biobank at the Department of Pathology, UZA, collaborates with surrounding hospitals for sample collection and contributes to the development of a virtual tumour biobank of the Belgian Cancer Registry.
Patricia Casbas-Hernandez
Ponce Health Sciences University- Ponce Research Institute, USA
Title: The Puerto Rico BioBank: The First Cancer Tissue Biobank at a US Hispanic-Serving Institution
Time : 15:40-16:05
Biography:
Patricia Casbas-Hernandez is a graduate from ‘La Universidad Complutense de Madrid’ (Spain) in Biochemistry, after which, she pursued her PhD in Molecular and Cellular Pathology in the University of North Carolina at Chapel Hill (NC, USA). Her interest in population based sciences led her to peruse post-doctoral training and a Masters in Public Health in Epidemiology at the same institution. She is currently a junior faculty member at the Ponce Health Sciences University (Ponce, PR) and co-leads the first Puerto Rico tissue BioBank.
Abstract:
There are 580 million people worldwide considered to be of Hispanic origin. In the US, Hispanics represent 17% of the population and its largest ethnic minority. Funded by NCI, Moffitt Cancer Center (Florida) and Ponce Health Sciences University (Puerto Rico) initiated a partnership that conducts high-quality research, training, and community-outreach focusing on cancer health disparities among Hispanics. A central component to this partnership is the Puerto Rico BioBank (PRBB), the first tumor bank capable of optimal collection, processing, and distribution of biospecimens derived from the island’s population. The PRBB has recruited cancer patients since 2008 through establishment of a functional laboratory infrastructure and collaborations with local hospitals. Eligible patients provide informed consent, agree to donate blood (for DNA) and tissues (fresh-frozen, paraffin embedded), and complete a comprehensive questionnaire related to clinical and lifestyle factors. Biological material is properly banked at the PRBB lab following HIPAA and standard operating procedures to ensure patient confidentiality and tissue quality. Currently, the PRBB has over 1,100 patients (65.8% females, 34.2% males) all of Hispanic origin. The most common banked tumors are breast, prostate, and lung, although other uncommon tumors are also collected. Pathological, demographic and clinical data is entered into a biobanking-management-system (BMS), which allows for up-to-date information and reporting. In conclusion, the patient samples banked in the PRBB are a unique resource that can support molecular and population studies. This Hispanic-specific biobank fosters collaborations among local and international researchers that facilitate the application of novel techniques to solving cancer health disparities among Hispanics.