9th Annual Conference on ^{Stem Cell and Regenerative Medicine} Sep 25-26, 2017 Berlin, Germany

Biography:

Karin Schuetze is a Biologist and expert in non-contact cell handling and label-free cell analysis using innovative photonic technologies. She did her PhD work at the Institute for Applied Physical Chemistry in Heidelberg and her Postdoc Research at the University of California in Berkeley where she assembled her first Optical Trap. In 1993 she and her husband founded the PALM company, which was successfully sold to Zeiss. In 2008, they both founded their second company, the CellTool GmbH, where they develop Raman microscope systems for biomedical applications, providing physicians and biologists with a label-free and non-invasive tool for cell analysis.

Abstract:

In the last years, the field of stem cell medicine has opened unforeseen therapeutic opportunities. In order to ensure functionality and safety of cell based therapies used for patient care, methods allowing reliable quality control during all steps of product development are essential. Raman Spectroscopy (RS) is a highly sensitive technology increasingly used for biomedical applications like cell identification, diffentiation and characterization, as well as quality control of cell based products as it reveals detailed information on the metabolic state of living single cells in a label-free and non-invasive way. Here, we demonstrate that RS is a valuable tool also in stem cell research and medicine. It helps to understand basic cellular processes, to ensure quality of cell based products and to improve current gold standard methods. We provide latest data from Raman analyses of single cells as well as of 3D cell culture and tissues, including investigations on retina tissue, insights on the influence of stem cell freezing and quality control of chondrocytes and blood products, showing that this non-invasive approach provides new insights on the whole metabolome of cells which help to improve and secure approaches in stem cell medicine.

Biography:

­­­Francisco Ruiz-Navarro is a Research Associate and R&D Director of the Austrian Society for Regenerative Medicine in Vienna, Austria where he focuses his research in stem cell therapies for neurological disorders. Before, he was performing research in the Cerebrovascular Department at the National Institute for Neurology and Neurosurgery in Mexico City with special interest in multicentric population studies with Hispanic stroke patients. He was a Research Assistant in the Brain Bank, Center for Research and Physiology, Department of Biology and Neuroscience at the Institute of Advanced Studies at National Polytechnic Institute in Mexico City. He obtained his Medical degree in the Anahuac University in Mexico City, became USMLE board certified in the United States of America and he got his Doctor of Medicine from the Medical University of Vienna.

Abstract:

After reviewing several current clinical guidelines, we found that less than 1% of all the recommendations for neurological diseases in the clinical guidelines are sustained by a level 1A of evidence (Systematic review of RCT with homogeneity, according to the Oxford Centre of Evidence). When reviewing the scientific evidence of stem cells treatment, we found that several stem cell therapies are based on the same or even stronger scientific evidence than the current medical treatment. This means accepted treatments and stem cell treatments nearly have the same level of evidence. Although both are far from level A, it is apparently enough for several clinicians and regulatory agencies to maintain a reluctant posture towards stem cell therapies but not towards other kinds of treatment. We propose that not just level 1A of evidence should be accepted as treatment, especially in cases where there are no options for patients besides experimental treatments. In this debate, the necessity of RCT or blinded trials arises, especially because SCs therapies are a procedure rather than medication and conducting clinical studies with “surgical placebos” is ethically controversial. There are analyses that describe how series of observational studies found results similar to those obtained from RCTs, which means that certain number of cohort or case-control studies would yield enough valuable information to bring treatments to the clinical practice, although we plan to wait for better-design RCT to confirm the results, if possible. We reviewed the most relevant literature concerning the use of stem cells to treat several neurological disorders. This literature review will provide key information on how stem cell’s interventions were performed with the analysis of the level of evidence they are providing. Drawing on the gathered information we then compared the scientific evidence that underlies the current treatment guidelines with the scientific evidence on currently available stem cells publications. As a result of this survey and analysis we conclude with a series of recommendations for the use of stem cells in clinical settings.

Biography:

MichaÅ‚ PikuÅ‚a is an Associate Professor at the Department of Clinical Immunology and Transplantology, Medical University of Gdansk. He has completed his MPharm from Medical University of Gdansk (Pharmacy Practice), MSc from University of Gdansk (Molecular Biology) and his PhD in Medical Biology from Medical University of Gdansk (Faculty in Medicine) in 2007. He is currently responsible for several projects focused on Regenerative Medicine and Experimental Immunology. He is also a Group Leader in a prestigious REGENNOVA consortium grant STRATEGMEDI (“Novel Technologies for Pharmacological Stimulation of Regeneration”).

Abstract:

Human adipose tissue constitutes a rich source of multipotent stem cells. Adipose-derived Stem Cells (ASCs) have been in the center of scientific and clinical attention, due to a wide range of possible implementations in tissue engineering, regenerative medicine and clinical immunology. These cells are characterized by a multipotent activity including high proliferative potential, secretion of trophic factors, direct interactions with immune cells and ability to differentiate into different kind of cells. Despite being extensively studied, phenotypic characterization of specific surface markers and details of differentiation potential need to be elucidated. The data obtained in our studies, based on flow cytometry technique, show that ASCs cultured in vitro express elevated levels of ASCs markers typical for mesenchymal stem cells (CD73, CD90, CD105, CD29). The variable level of expression was observed for CD31 and CD34, what seems to be a donor-to-donor variability. The observed markers expression did not affect the differentiation capacity of ASCs into chondrocytes, adipocytes and osteocytes. Additionally, our results show that the long term in vitro culture of ASCs has no impact on the surface expression markers pattern. According to our observations, no significant differences in the surface markers level and differentiation potential were noted between the cells obtained from oncological patients or from plastic surgery procedures. Summarizing, our studies reveal that ASCs are a promising tool in postoperative tissue reconstruction in oncological patients due to their high regenerative potential. However, further studies are required to provide more insight into the therapeutic potential of ASCs in postoperative management of patients undergoing oncology surgery.

Biography:

Vassilis Katsares is a Biologist with a PhD on Populational Molecular Genetics. He completed three Postdoctoral Fellowships. He also served as a lecturer for undergraduate and postgraduate courses. He had worked as Technical Sales Support for immunology analyzers and afterwards he was the Laboratory Director at a biopharmaceutical company. His main research interests cover many aspects of Molecular Genetics, Cellular Therapy projects, and bioethics. He has authored 21 scientific papers and 36 announcements at national and international conferences. He is a referee in various journals. He is member in many international Scientific Organisation and Societies. His CV is included in “Who is Who in the World”, and in “Who is Who in Science and Engineering”.

Abstract:

Stem cells play a significant role in the novel branch of modern Biology and Medicine, known as Regenerative Medicine. Due to their regenerative ability, stem cells are looked at as a promising tool for improving infertility treatments in women and men. It has been shown that fully mature egg cells, as well as sperm cells can be grown in the lab, raising hope for new infertility treatments. There has been also evidence that eggs could be fertilised, implanted into a surrogate female and go on to produce live offspring. All the obtained offspring grew up normally without evidence of premature death. Artificially created sperm and eggs could spell the end of infertility at the near future. Stem cells can potentially be developed into any tissue in the human body. Reproductive scientists think it is only a matter of time before the technique is used as an alternative IVF treatment. Nowadays, there is also the new trend of ovarian rejuvenation, using stem cells growth factors to improve the ovulation of older ovaries. However, for all these techniques to become a clinical reality, many potential risks and ethical concerns about the use of stem cells must first be addressed. If the treatment works, it could be extended to even older post-menopausal women. Should the law review the ceiling on how old a woman can be to have this treatment since there is no law stopping a man from having a child late in life? All this research may alter our understanding of male and female infertility and provide solutions to IVF treatments. Though a long way off, if ever, it would be an incredible change in how we view and treat infertility.

Biography:

Joseph Choukroun completed MD from University of Montpellier, France 1979 and is a Specialist in General Surgery, Anesthesiology from the same university. He is also a Specialist in Pain Management from University of Strasbourg France. He is the Owner of Private Pain Clinic, Nice France. He is the President of SYFAC, international symposium on growth factors. He is the Inventor of the PRF techniques: L-PRF, A-PRF and i-PRF. He is a Researcher working in Form Lab at University of Frankfurt. He is the author of several scientific publications and is recognized as an International Speaker.

Abstract:

Regenerative therapy with stem cells has gained tremendous momentum over the past decade as a modality geared towards markedly improving wound healing of various tissues by utilizing undifferentiated autologous host cells. While stem cells may be isolated from various locations in the human body, more recently it has been shown that low levels of mesenchymal stem cells also exist circulating within peripheral blood. Platelet rich fibrin (PRF) is a regenerative modality that utilizes peripheral blood + centrifugation protocols without the use of anti-coagulants to create a three-dimensional tissue engineering scaffold containing both growth factors and autologous cells. Very recently, it has been shown that modifications to centrifugation speed and time following recently developed concepts (the low-speed centrifugation concept or LSCC) resulted in a marked increase in host cells and growth factors. Within these scaffold constructs, mesenchymal stems cells were also found following collection with this relatively painless and low-cost modality. The objective of the present talk will be to present recent modifications to centrifugation speed and time to optimize stem cell quantities within PRF. Thereafter, the biological data supporting their numbers, as well as their potential for clinical applications will be presented with data coming from many fields of medicine including for the regeneration of osteoarthritic knees, dental regenerative medicine, orthopedic grafting, and for facial esthetics.

Biography:

Simon Haas is Group Leader at the Heidelberg Institute for Stem Cell Technology and Experimental Medicine and the German Cancer Research Center. He is an expert in hematopoiesis, with a special focus on stem cell biology. His research centers around the question how hematopoietic stem cells make complex lineage decisions to produce the variety of blood and immune cells.

Abstract:

Multipotent hematopoietic stem cells are responsible for the livelong production of all blood and immune cells. In the classical model of hematopoiesis, blood formation is believed to occur through stepwise progression of hematopoietic stem cells following a tree-like hierarchy of oligo-, bi- and unipotent progenitors. However, this model is based on experimental approaches unable to describe how individual HSCs and their progeny enter lineage commitment during steady-state hematopoiesis. To establish a comprehensive model of human hematopoiesis, we have developed single-cell approaches that integrate single-cell RNA-sequencing with flow cytometric and functional lineage potency data. This allows us to reconstruct developmental trajectories and to gain a detailed view on lineage commitment of individual HSCs into all major branches of human haematopoiesis. We found that individual HSCs do not pass through discrete intermediate progenitor cell stages. In contrast, HSC lineage commitment occurs in a gradual manner best described by a continuous Waddington landscape with initially flat but progressively deepening valleys. Our data determine a detailed model of developmental trajectories within this landscape and demonstrates that distinct gene expression modules operate in a combinatorial manner to control stemness, early lineage priming and the subsequent progression into all major branches of hematopoiesis. These results establish the concept of a developmental continuum, which can replace the differentiation tree as a comprehensive model of human steady-state hematopoiesis and provide a basis for the understanding of hematopoietic malignancies.

Biography:

Hany E. S. Marei is the Project Manager of Biomedical Research Center, Qatar University since May 2015. His research interest lies in the field of neural stem cells, induced pluripotent stem cells, genomics, transcriptomics and epigenomics of Neurological diseases/disorders. He is the editorial board member/reviewer for several international journals. He served as a member for promotion committee for Professors at the Supreme Council of Universities Egypt from 2012-2015.

Abstract:

Neural stem cells (NSCs) are multipotent self-renewing cells that could be used in cellular-based therapy for a wide variety of neurodegenerative diseases. Here, to assess the ability of carbon nanotubes (CNTs) to enhance the therapeutic potential of human olfactory bulb neural stem (OBNSCs) for restoring cognitive deficits and neurodegenerative lesions, we co-engrafted CNTs and human OBNSCs in trimethyltin(TMT) -neurodegeneration rat model. The present study revealed that engrafted human OBNSCS-CNTs restored cognitive deficits, and neurodegenerative changes associated with TMT-induced rat neurodegeneration model. Moreover, the CNTs seemed to provide a support for engrafted OBNSCs, with increasing their tendency to differentiate into neurons rather than into glial cells.

Biography:

Tamara Lah Turnšek has completed her graduation (organic chemistry), magisterium (biochemistry) and doctorate (biochemistry), she worked at the Department of Biochemistry and Molecular Biology at the Jožef Stefan Institute and for quite a while likewise at the Clinical Laboratory of the Paediatric Clinic in Ljubljana. Since 1996, she has been Director of the National Institute of Biology, where in the vicinity of 1997 and 2004 she additionally filled in as Head of the Department of Genetic Toxicology and Cancer Biology.

Abstract:

The most aggressive brain tumour glioblastoma multiforme is characterized by aggressive GBM cell infiltration into surrounding brain parenchyma. We hypothesise that this invasion process is supported by mesenchymal stem cells, comprising GBM microenvironment. MSCs are recruited from bone marrow or endogenous brain tissue to the GBM tumour, but their interactions with GBM cells are still poorly understood. To elucidate the direct interaction between bone marrow-derived MSCs and two distinct GBM cell lines, U87 and U373, we tested cells’ invasion in vitro, as well as in vivo, using zebrafish embryo model. Since proteases are crucial for GBM cell invasion, we focused on their role in invasion of cells in MSC/GBM direct co-cultures by analysing their expression at gene and protein levels and by applying selective protease inhibitors in the 3D-invasion model in vitro. We demonstrated that the effect of MSC/GBM cellular cross-talk on GBM cell invasion is GBM cell type specific. Namely, MSCs decreased the invasion of U87 cells, whereas they increased the invasion of U373 cells in vitro and in vivo. In contrast, both GBM cell lines increased the invasiveness of MSCs upon direct interactions. Moreover, we observed that increased U373 cell invasion in co-cultures correlated with increased expression of cathepsin B, calpain1, uPA/uPAR, MMP-9 and -14, all involved in the protease signalling cascade in GBM cells, leading to increased invasion via extracellular matrix degradation. Using selective inhibitors, we confirmed involvement of cathepsin B, MMP-9 and -14 in MSC-enhanced invasion of U373 cells. By contrast, decreased invasion of U87 upon co-culturing seemed to be independent of these proteases, implicating that the MSC regulatory potential in MSC/GBM co-cultures is dependent on GBM phenotype. Finally, we identified the genes, associated with cell response to TGF-ß that were differentially expressed in U87 vs. U373 cells that could explain different response of these cell lines to MSCs. Taken together, our findings are the first to suggest that the response of GBM cells to MSCs depends on the cancer cell’s genetic subtype. This notion may be generalized to other types of stromal cells as well as to other tumours.

Biography:

Walter Birchmeier is a graduate in biology of Zürich University in 1973. After postdoctoral years at Cornell University Ithaca, the Biocenter Basel, and the University of California at San Diego, he became laboratory head at the Max-Planck-Institute Tübingen in 1982 and full Professor at the University of Essen Medical School in 1988. He joined the Max-Delbrück-Center for Molecular Medicine in Berlin in 1993. He was Director of the Max-Delbrück-Center from 04/2004 to 12/2008 and is Professor at the Charité/Humboldt University Berlin. His major research interests have been the role of cell adhesion and signal transduction in development and tumor progression.

Abstract:

Head and neck squamous cell carcinomas (HNSCCs) of the upper airways, which also include salivary gland cancers, are the fifth frequent human malignancy. The most important risk factors for HNSCCs are smoking, excess alcohol consumption and infection by high-risk human papillomaviruses. Patients with advanced tumors exhibit high mortality due to lack of effective molecular therapies. We found in a mouse model of salivary glands squamous cell carcinoma with conditional beta-catenin gain-of-function mutation that a histone modifier, which induces H3K4me3, acts downstream of Wnt/beta-catenin signaling. Conditional ablation of the histone modifier gene prevented tumor formation, reduced proliferation and induced apoptosis. ChIP-seq revealed a genome-wide increase in the active histone mark H3K4me3 and chromatin opening in cancer stem cells (CSCs). Mutations by CRISPR/Cas9 of the histone modifier at the β-catenin-, Menin-, Brd4-, and Wdr5-binding and Set-H3K4me3 enzymatic sites in mice strongly reduced CSC self-renewal. Pharmacological interference that disrupted these interactions also strongly reduced the self-renewal of mouse and human CSCs. We thus identified an essential downstream layer downstream of Wnt/beta-catenin, which are H3K4me3 and opening of chromatin that are essential in HNSCC formation. Targeting the histone modifier and its interactions with small interfering molecules allow promising new therapies for head and neck squamous cell carcinomas.

Biography:

Zhengping Liu is an Obstetrician. He completed his MD from the Second Military Medical University in 1993. Currently, he is a Professor and Director of the Department of Obstetrics, Southern Medical University affiliated Maternal & Child Health Hospital of Foshan, and the Deputy Director of the Foshan Institute of Fetal Medicine. His research interests include placenta previa and abnormally invasive placenta, fetal in utero treatment, and regenerative medicine and stem cell clinical therapy. More than 30 articles reflect his research in reputed journals and has been serving as an Editorial Board Member of repute. He is the pioneer of fetal surgery therapy in utero in China, and has extensive collaborations with The Children’s Hospital of Philadelphia. So far, he already performed dozens of clinical participants with stem cells during the past year.

Abstract:

Ceasarean delivery has already become a very common way of delivery around the world, especially in low-income countries. Hypertrophic scars and wound infections have affected younger mothers and frustrated obstetricians for a long time. Previous studies have demonstrated that MSCs are involved in enhancing diabetic wound healing. Therefore, this study is designed to investigate the safety and efficacy of using MSCs in the treatment for the caesarean section skin scars. This trial is a prospective, randomized, double-blind, placebo-controlled, single-center trial with three parallel groups. Eligible participants are randomly allocated to placebo, low-dose (3×10⁶ cells transdermal hydrogel MSCs) or high-dose group (6×10⁶ cells transdermal hydrogel MSCs), once a day for consecutive six days. Study duration is 6 months. The primary outcome of this trial is to evaluate the change of Vancouver scar scale during the 6 months. Adverse events, including severe and slight signs and symptoms, are documented in case report form. The study is conducted at the Department of Obstetric of Southern Medical University Affiliated Maternal & Child Health Hospital of Foshan. The first participant was recruited on September 14^th, 2016. We hope to complete enrolment for the trial by September 2017 with all 6-month follow-up data expected by March 2018. This trial is the first investigation of the potential for therapeutic use of MSCs for the management of women’s skin scar after cesarean delivery. The results will give us an effective therapeutic strategy to combat caesarean section skin scars, even in the uterine scar.

Biography:

Ganapathi Bhat Mugulthimoole is Senior Consultant Medical Oncologist & Stem cell transplant Physician at Jaslok Hospital & Research Centre Since 2006. He gained specialized training in stem cell transplantation as part of the ESH-EBMT (2007), 2011(Labaule, France) and ICAS training program (2009) from ULM University, Germany. He is also a member of academic organizations namely ESMO, IELSHG, EHA, Asia Pacific Bone Marrow Transplant and an Affiliate of American Association for Cancer Research and BITs Congress Tank. He served on the board of teaching faculty (Kuwait) for MRCP (Pathology & Haematology). He is also editorial member of various international scientific Journals.

Abstract:

Allogeneic hematopoietic stem cell transplantation (HSCT) is cellular immunotherapy in the true sense for the treatment of a number of benign and malignant disorders with a curative intent. Delays in immune reconstitution following HSCT considerably limit the positive outcomes and increase the risk for infection and disease relapse in the transplant recipient. Ways to measure and manipulate immune recovery following HSCT are emerging and their success depends directly upon an enhanced understanding for the underlying mechanisms responsible for reconstituted immunity and haematopoiesis. Research in transplant immunology has made considerable efforts in understanding the role of immune effector cells in HSCT. Understanding the transplant immunobiology is fundamental to elucidate the immunological process involved in engraftment, immunotolerance, immune reconstitution, and donor and host reactions such as graft-versus-host disease (GVHD), graft-versus-leukaemia (GVL) effect, graft rejections and reinstatement of hematopoietic and immunological function to prevent transplant-related opportunistic infections. Although much of our understanding of immunobiology in allogeneic HSCT is from research in preclinical models, the findings can however be correlated with clinical observations in transplant recipients. Meanwhile, several pharmacological and cellular therapeutic interventions have been shown impact on immunobiology and influence the outcome. As our knowledge continues to evolve in understanding immunobiology, so will our ability to decrease disease- and transplant-related morbidity and mortality in the setting of HSCT.

Biography:

Asuman SunguroÄŸlu is Professor at Medical Faculty of Ankara University. She graduated from Hacettepe University, Faculty of Science, Department of Biology, Ankara, Turkey. She got the Master’s Degree in “Tumour Immunology” in 1987 from Medical Faculty of Hacettepe University. She received her PhD degree in “Medical Biology” from Ankara University, Medical Faculty in 1992. Her research focuses on cancer and leukaemia cytogenetics in humans. In order to specialise and gain knowledge and practice, she was assigned in the University of Welsh, Medical Faculty in Cardiff for 6 months in 1996. She was named Associate Professor in 1997 and Professor in 2003. She has been Chair of Medical Biology Department since 2003. She is the member of Advisory Committee of the Scientific and Technological Research Council of Turkey (TUBÄ°TAK). She received several grants, awards and fellowships from organizations such as Turkish Heart Association, Ankara University and TUBÄ°TAK for her research projects. Her current research interests in molecular mechanisms of cancer stem cells, understanding the molecular basis of Infertility, Monoclonal Antibody production and immunotherapies for cancer.

Abstract:

Background/Aim: Glioblastoma, known as Glioblastoma Multiforme (GBM), is the most common and aggressive type of brain tumours in adults and contains self-renewing,  tumorigenic cancer stem cells (CSCs) that can account for tumour initiation and acquisition of resistance to the given anti-GBM therapies. The neural stem cell marker CD133, known as prominin-1, has been widely used as a CSCs marker in GBM. Although, there is some controversy regarding tumour-initiating properties of CD133+ and CD133- GBM cells, a growing number of studies have revealed GBM initiating stem cell capability of CD133+ cells. Elucidation of the molecular characterization of GBM CSCs is essential for the development of novel targeted therapeutics for GBM. Therefore, we aimed to determine the expression levels of other potential CSC markers in CD133+ GBM CSCs.

Materials and Methods: Primary GBM cells were isolated from freshly obtained GBM tissue samples derived from ten individual patients. These cells were cultured with DMEM with high glucose including 1% Penicillin-Streptomycin and 10% fetal bovine serum. Then, CD133+ and CD133- cells were separated by MACS (Miltenyi) method from those GBM primary cells. The CD133+ selected and CD133- cell populations were collected in different tubes. Following RNA isolation from CD133+ and CD133- cells, cDNA synthesis was performed. mRNA expression levels of 88 genes were detected by Real Time Cancer Stem Cell PCR Array (Bio-Rad). The Student T test was used to identify statistically significant differences between groups. Differences were accepted to be statistically significant at p<0.05. Confocal microscopy was performed to examine the localization of CD38 and CD24 proteins in CD133+ GBM CSCs.

Results: Based on the results of PCR Array, we found that the mRNA levels of ABCG2, ALCAM, CD24, CD38, CD4, DDR1, EGF, ENG, ETFA, FGFR2, FLOT2, FZD7, GSK3B, ID1, IKBKB, ITGA2, ITGA4, ITGA6, ITGB1, JAG1, MAML1, MUC1, MYCN, NFKB1, NOTCH2, PLAT, PLAUR, POU5F1 and BMP7 were statistically different in CD133+ GBM CSCs when compared to expression of those in CD133- cells. Furthermore, we investigated protein levels of CD38 and CD24 in both CD133+ and CD133- cells and observed that protein expression of both CD38 and CD24 was more prominent in CD133+ GBM CSCs than in CD133- cells.

Conclusion: Our results suggest that in addition to the presence of CD133 expression GBM initiating cells have also the expression of different genes involved in distinct survival pathways, indicating tracing these possible candidates could be useful for characterization of CD133+ GBM stem cells. This research has been supported by The Scientific and Technological Research Council of Turkey (No: 114S189).

Biography:

Dilara Akcora Yildiz is an Assistant Professor at Biology Department, Mehmet Akif Ersoy University. She has been the Vice Director at the Institute of Science and Technology since 2016. She graduated from the Biology Department, Faculty of Science, Ege University, Turkey in 2004. She received her master’s degree from the Medical Biology Department, Faculty of Medicine, Ankara University, Turkey in 2007 and studied the effect of T315I, E255K and M351T mutations in imatinib resistance in chronic myeloid leukaemia patients. In the same year she was awarded with a Postgraduate Education Scholarship in Australia by the Ministry of National Education of Turkey (MEB) (2008-2012). She then earned her Ph.D. degree in intestinal stem cell biology at Department of Pathology at The University of Melbourne in 2012 under the supervision of Prof. Dr. Robert G. RAMSAY. During her doctoral studies, she characterized the role of colony stimulating factor 1 receptor-ligand pair (Cfms/CSF1) and granulocyte macrophage colony-stimulating factor (GM-CSF) in intestinal biology. She was the principal investigator of a research project titled as ‘The effect of WRN and MGMT proteins which play a role in  DNA repair on drug resistance occured in Multiple Myeloma disease’ and supported by The Scientific And Technological Research Council Of Turkey (TUBITAK - 3501 National Young Investigator Career Development Program). She is currently working as an investigator in other projects focused on brain tumors, cancer stem cells and antibody production. Dr. Akcora Yildiz was a participant at 9th HOPE Meeting with Nobel Laureates in 2017. Her research interests include stem cell biomarkers, DNA repair mechanisms, apoptosis and autophagy signaling in cancer biology.

Abstract:

Glioblastoma Multiforme (GBM), is the most prevalent and aggressive type of primary brain tumour with a median survival of only 15 months due to recurrence of tumour after surgical resection and acquisition of resistance to radiotherapy or chemotherapy. Temozolomide (TMZ) an oral alkylating agent leading to the occurrence of DNA damage has been still used for GBM treatment. Other than TMZ, Bortezomib (BZ), currently in clinical use for the treatment of myeloma by achieving proteasome inhibition, has been revealed to induce apoptosis and growth inhibition in GBM cells. Our purpose was to examine the role of potential cancer stem cell markers including CD133, CD38, CD24, CD70 and DR6 in cell survival after either TMZ or bortezomib treatment. U118 GBM cell line and other GBM cell lines including U87, U138 and T98 were incubated with TMZ and BZ for 48 hr, respectively.  Real Time Cancer Stem Cell, Integrin, Apoptosis and Cell Adhesion PCR Arrays (Bio-Rad) were performed in U118 cells treated with TMZ for 48 hr. Flow cytometry assay was used to determine the protein amounts of the genes of interest after BZ treatment. Treatment with TMZ led to an increase in mRNA expression of CD38 and CD24 but not in CD70 and DR6. BZ decreased the expression of CD133 and CD38, whereas CD24 expression was found to be increased in a dose-dependent manner in all GBM cell lines. Furthermore, CD70 protein expression was elevated, while DR6 protein expression was reduced with the increase of the dose of BZ. Our results suggest that CD24 seems to be involved in GBM cell survival after either TMZ or BZ treatment, indicating inhibition of its expression might benefit to overcome chemo resistance. This research has been supported by The Scientific and Technological Research Council of Turkey (No: 114S189).

Biography:

Viacheslav M Mikhailov received education in Medicine from Mechnikoff’s Medical University during 1958-1964 years. He pursued Postgraduation from Embryological Department of Institute of Experimental Medicine (1958-64) and PhD degree (1970). His PhD thesis entitled: “The analyses of mechanisms of pathogenic action of immune antirenal antisera for mammalian embryogenesis”. He was awarded DS degree for thesis: “Life Cycle of Decidual Cells”, by title Leader Research Fellow and by Professor title in 1998 and 2003 respectively. He is the Head of Cell Populations Genetic Group of Institute of Cytology RAS.

Abstract:

Cell interactions between uterus and foetuses influence for success of pregnancy and of fetal development in mammals. In case of human and rodents the embryos and foetuses are surrounded by decidua cells (DC) that form decidua membrane. Without DC blastocyst development of experimental animals stops at the initial stages of gastrulation and doesn’t go beyond somites formation. DC participate in the trophic relationships between mother and foetus, their specific functions include the prevention of the development of inflammation in the endometrium and the regulation of immunological conflict between mother and foetus. There are also results that single intravenous transplantation of pregnant rats Percoll derived mononuclear bone marrow cells (BMC) suspension of 4-5, 7-9 or 11-12 pregnant days to rats with the same date of pregnancy influences for survival and weight of foetuses of 18th of pregnancy. After BMC, intravenous transplantation during 4-5 days of pregnancy the weight was preserved but survival of foetuses was decreased. The weight of 18th day foetuses after transplantation during pregnant 7-9 days (835 ± 15 mg) was significant increase in comparison with weight of normal and control foetuses (745 ± 11). The survival of foetuses was also preserved. In case of intravenous transplantation during placentation at 11-12 days of pregnancy the weight of foetuses was decreased, the weight of placentas was increased and survival of foetuses was disturbed. The retardation of foetuses growth after BMC transplantation during placenta formation (587 ± 5 mg) at pregnant 11-12 days may be explained by cytotoxic action of uNK cells for embryo. At the same time, the sub-dermal BMC transplantation at 11 and 13 pregnant days stopped the embryotoxic action of uNK cells, increased the weight of foetuses and preserved the survival of embryos.

Biography:

Helen McGettrick completed her PhD in 2006, followed by 3 Postdoctoral Research fellowships at the University of Birmingham (UK). She was appointed as a University Fellow in Inflammation Biology in 2011, and a year later successfully won a five-year Arthritis Research UK Career Development Fellowship. She was recently awarded the prestigious Garrod Prize by the British Society for Rheumatology in 2016. She is an Honorary Lecturer at the University of Glasgow and Newcastle University (UK). She has more than 30 published articles with about 760 citations, and has filed 3 patents and received funding from Wellcome Trust, Pfizer and British Heart Foundation.

Abstract:

Chronic inflammation is associated with formation of ectopic fat deposits that might represent damage-induced aberrant mesenchymal stem cell (MSC) differentiation. Such deposits are associated with increased levels of inflammatory infiltrate and poor prognosis. Here we tested the hypothesis that differentiation from MSC to adipocytes in inflamed tissue might contribute to chronicity through loss of immunomodulatory function. We assessed the effects of adipogenic differentiation of MSC from bone marrow or adipose tissue on their capacity to regulate neutrophil recruitment by endothelial cells and compared the differentiated cells to primary adipocytes from adipose tissue. Bone marrow derived MSC were immunosuppressive, inhibiting neutrophil recruitment to TNFα-treated EC, but MSC derived adipocytes were no longer able to suppress neutrophil adhesion. Changes in IL-6 and TGFβ1 signaling appeared critical for the loss of the immunosuppressive phenotype. In contrast, native stromal cells, adipocytes derived from them and mature adipocytes from adipose tissue were all immuno-protective. Thus, disruption of normal tissue stroma homeostasis, as occurs in chronic inflammatory diseases, might drive abnormal adipogenesis which adversely influences the behavior of MSC and contributes to pathogenic recruitment of leukocytes. Interestingly, stromal cells programmed in native fat tissue retain an immuno-protective phenotype.

Biography:

Jasbir Rattu is a Consultant Industrial Pharmacist EU qualified person focussed on innovative regenerative medicines and advanced therapy medicinal products for unmet medical need.

Abstract:

IQ= QRM x QbD x QMS. Quality risk management is an enabling process that supports the product life cycle and is an integral part of an organisation’s pharmaceutical quality system. The ultimate goal of the QRM process is to bring focus and effort to the issues that impart the highest risk to product quality and/or patient safety. Therefore, as QRM is ultimately linked to the protection of the patient, a modern PQS cannot truly function properly in the absence of an effective, integrated QRM system. QRM is a systematic process for the assessment, control, communication and review of risks to the quality of the medicinal product. It can be applied both proactively and retrospectively. Therefore, the objective is not just to identify risk, but to provide information to help make better and informed decisions to mitigate and reduce risk, thus improving the process. Hence, it is critical to understand and effectively and optimally use the most appropriate risk management tools and approaches. The consistent and effective analysis of risks associated with manufacturing processes and quality systems typically leads to more robust decisions reduces uncertainty and leads to greater confidence in outcomes. These QRM elements must be embedded throughout the organisation through policies, procedures and reinforced via effective training. Quality by design requires a deep understanding of the product and process science which impacts quality and hence performance. Quality Management System is the set of systems which ensure high quality is continuously improved e.g. audit systems, change control systems, deviation management systems etc. Continuous improvement of QRM multiplied by excellent QbD understanding and again by reliable Quality Management Systems provides a powerful optimised integrated quality system. This presentation discusses best practice including my experience of successfully implementing integrated quality systems in high risk areas such as regenerative medicine and other areas of the biopharmaceutical industry including applications to cutting edge advanced therapy medicinal products e.g. cell and gene therapies such as CAR-T cells in immuno-oncology to the tissue engineering of organs such as the eye and stem cell-based therapies for treating optic neuropathies such as glaucoma.

Biography:

Luca Gentile graduated in Developmental Biology in 2000 and received his PhD in Bioengineering and Bioinformatics at the University of Pavia (ITA). In 2003, he was invited to the Center for Animal Transgenesis and Germ Cell Research (Kenneth Square, PA, USA) for studying reprogramming in single reconstructed mouse embryos. In 2004, he joined the Max Planck Institute for Molecular Biomedicine (Münster, GER), working on the onset of cellular reprogramming. In 2010, he was appointed Head of the Planarian Stem Cell Laboratory (Münster, GER), working on the conserved mechanisms of pluripotency. In the same year, he organized the first International Meeting on Planarian Biology, which in 2018 reaches its 4^th edition (Madison, WI, USA). Since March 2015, he leads the Pluripotency & Regeneration Group at the Fraunhofer Institute for Biomedical Engineering (Sulzbach, GER). His subjects of investigation are the derivation of human cardiac organoids and the functional characterization of stem cell subpopulations in planarian S. mediterranea.

Abstract:

Over the last 60 years, 14% of all drugs were withdrawn post-marketing owing to their cardiotoxic effects. Cardiovascular diseases are responsible for nearly half of all deaths in Europe. This is largely due to the poor regeneration capability of the human heart, which culminates in scar tissue formation and reduced function. Although advanced medical procedures increased the survival rate, cell-based therapies fall short in delivering a functional engraftment, and many patients progress towards end-stage heart failure. We used 3D printing and ultra-high viscosity alginate to produce a hydrogel scaffold called the Heart-patch, which could sustain cardiac muscle identity as a functional unit over long time in culture. Human induced pluripotent stem cell-derived cardiomyocytes cultured on the Heartpatch are closer to adult CMs that those cultured on traditional plastic, making the Heartpatch a physiologically relevant model of the heart. Biocompatible UHV alginate is 3D printed using a novel, viscosity-independent, printing method. It allows both patterning the scaffold’s surface and tuning its stiffness. Additionally, surface modifications can be applied with click-it chemistry, allowing a specific ECM protein composition that mimics the cardiac niche. Cardiomyocytes on the Heartpatch can be cultured for longer periods (3-4 months) than their counterparts on cell culture plastic, exhibiting self-sustained macro contractions visible to the naked eye. Gene expression profiling and immunocytochemistry analysis indicate increased cardiomyocyte maturation, enhanced cell-cell interactions and a high degree of both electrophysiological coupling and cytoskeletal maturation. As a result, force transduction resembles more closely the in vivo situation, as also supported by a novel optical-based analysis of the self-sustained depolarization. On the Heart-patch, hiPSC-CMs acquire an additional degree of freedom. As an individual working unit, the Heartpatch is an enhanced cardiac muscle model that enables disease modeling and high-throughput compound screening, with potential future applications in regenerative medicine.

Biography:

Jiamin Teng is a Professor in the Department of Pathology and Translational Pathobiology, Louisiana State University Health Sciences Center in Shreveport (LSUHSC), USA. She completed Master’s Degree from Tongji Medical University in Wuhan, China and her PhD in Nihon University School of Medicine in Tokyo, Japan. Her research focuses on glomerulosclerosis of kidney damage and the stem cell regeneration, which can be translated to diabetic nephropathy due to similarities in pathogentic mechanisms involved. Her laboratory has developed unique experimental models such as in vitro live cell micromanipulation model, in vitro 6 dimensional live cell observation models, ex vivo kidney perfusion model, and in vivo model for the kidney glomeruli damage and reparation. These models have well served for the purpose of kidney pathogenic and regeneration.

Abstract:

Many investigators have supported the idea that mesenchymal stem cells (MSCs) participate in the process of repair / regeneration exclusively by providing paracrine factors that enabled the process.  Using a model of mesangial damage induced by glomerulopathic immunoglobulin light chains (G-LCs) and then repaired or regenerated by MSCs in AL-amyloidosis, the role of MSCs in the process was investigated. In-vitro and ex-vivo experimental platforms were used to address the issue. The in vitro 6 dimensional (6D) live cell imaging system was used to observe the damage of mesangial cells (MCs) and the alteration of the mesangial matrix incubated with G-LCs. In the ex vivo model, G-LCs were perfused through the renal artery. The respective lesions were reproduced in both platforms. Then, tagged MSCs were introduced. Immunofluorescence, immunohistochemistry and electron microscopy were used to evaluate samples obtained at different time frames. Stains for smoothelin, muscle specific actin, smooth muscle actin, CD29, and 68 were used to monitor phenotypic transformation of MSCs in the process of regeneration. Our results showed that MSCs initially transformed from an undifferentiated to a macrophage phenotype to clear the damaged mesangial areas where transformed MSCs phagocytosing cellular debris resulting from apoptotic mesangial cells and damaged matrix elements of amyloid fibrils. Following the cleaning process, MSCs acquired morphologic and immunophenotypic characteristics of MCs as they proceeded to lay down new mesangial matrix. MSCs manifest great plasticity as they proceed to repair the damaged mesangium in both our models. The fact that they transform to a macrophage phenotype followed by transformation to MCs allows them to perform different crucial functions during the process of regeneration. The restored mesangium is possible as new MCs derived from MSCs are able to reproduce the normal mesangium.

Biography:

Sharmila Fagoonee is a Researcher at the Institute of Biostructure and Bioimaging of the Italian National Research Council at Molecular Biotechnology Center in Turin, Italy. After her BSc (hons) in Biology at Mauritius and MSc in Cellular Biology at Bordeaux, France, she got her PhD in Cell and Molecular Biology at the University of Turin. She also holds a MSc in Molecular Biotechnology. During her PhD, she worked on heme and iron metabolism in mouse models of liver diseases. After a period at BIDMC, Harvard, Boston, she shifted to study the differentiative capacity of mouse pluripotent spermatogonial stem cells in vitro and in vivo. Using bioinformatics analysis, she identified new pluripotency- and stemness-related genes in pluripotent stem cells. She is currently interested in human stem cells-based therapy for liver diseases and in studying the mechanisms leading to and in developing new biomarkers for liver fibrosis.

Abstract:

Adult stem cells have gained in attractiveness over embryonic stem cells for liver cell therapy due to their origin, multipotentiality, and the possibility of autologous transplantation. In the last two decades, great strides have been made in spermatogonial stem cell research showing the potentiality of these stem cells in organ regeneration and in the field of regenerative medicine. Spermatogonial stem cells derived from post-natal or adult mice are capable of differentiating into spermatozoa in vivo. A spontaneous embryonic stem cell-like conversion is also observed in vitro in long-term cultures. These pluripotent spermatogonial stem cells can be induced to differentiate into cells of the three germ layers without ethical constraints. We have recently shown that functional hepatocytes can be derived from mouse pluripotent spermatogonial stem cells for in vitro and in vivo use. These cells can thus provide a very promising source of hepatocyte-like cells for the regeneration of damaged tissue and for restoring liver functionality.

Biography:

Christopher A Bradley completed his PhD in Biochemistry & Molecular Biology at Louisiana State University Health Sciences Center in Shreveport. His Postdoctoral research was in the fields of eukaryotic protein synthesis, proteomics, cancer, osteoarthritis, and virology. Industry experience in detergent chemistry and nutraceuticals preceded his current position in regenerative medicine as Product Development Director for Lattice Biologics, a manufacturer of stem cell and tissue allografts.

Abstract:

The extracellular matrix consists of structural and functional molecules secreted by the resident cells. The 3-dimensional organization and composition of an ECM is distinctive for each tissue type. Solubilized gel-forming ECM could potentially be used to develop products such as 3D culture substrates, growth factor delivery vehicles, or scaffolds that promote tissue regeneration. One goal of ours is to design a 100% allograft custom-fabricated human ECM-derived 3D microenvironment that mimics the biological properties of native ECM. Toward this goal, we evaluated the biochemical and functional characteristics of ECM isolated from human placenta, adipose, and cadaveric muscle tissues. In addition, we compared the properties of muscle-derived ECM isolated from young versus old donors. Proteomic analysis showed the presence of proteins involved in matrix remodeling, angiogenesis, cell growth and differentiation. ECM protein variability largely depended on tissue type and donor age. Regardless of tissue origin, ECM-based 3D gels affected cell morphology by reducing cell spreading. Differentiation of adipose-derived stem cells subjected to ECM sourced from various tissues showed no effect on adipogenic ADSC differentiation. Contrastingly, we found that the presence of young muscle ECM augmented chondrogenic ADSC differentiation, while old muscle ECM favored osteogenic ADSC differentiation. In summary, tissue- and age-related properties of ECM should be considered in developing 3D culture models for basic and translational research.

Biography:

Andriana Margariti has developed significant expertise in stem cell biology, with particular emphasis on cell reprogramming, chromatin remodelling, cell signalling and endothelial cell biology. Her research program is based on the remarkable idea of direct reprogramming which will allow her to realize fundamental principles of cell reprogramming and establish homogeneous populations of endothelial cells. Development of fast and robust new methodologies that produce well-characterized, homogenous, clinical-grade cells suitable for tissue repair/re-modelling would have great utility. Her research team has generated patient-specific iPS cells and PiPS cells and they now have exciting novel data and they are working towards to develop a remarkable new highly efficient strategy of cell reprogramming. Importantly, the potential of the reprogrammed endothelial cells to enhance angiogenesis and neovascularisation and to promote perfusion of ischemic tissue will be established. Her research is demonstrating the therapeutic potential of “re-born” reprogrammed endothelial cells which would have transforming consequences for regenerative and personalized medicine.

Abstract:

The capability to derive endothelial cell (ECs) from induced Pluripotent Stem (iPS) cells holds huge therapeutic potential for diabetes. This study elucidates the precise role of the RNA-binding protein Quaking isoform 5 (QKI-5) during EC differentiation from both mouse and human iPS cells and dissects how RNA-binding proteins can improve differentiation efficiency towards cell therapy for important vascular diseases such as diabetes. iPS cells represent an attractive cellular approach for regenerative medicine today since they can be used to generate patient-specific therapeutic cells towards autologous cell therapy. In this study, using the model of iPS cells differentiation towards ECs, the QKI-5 was found to be an important regulator of STAT3 stabilisation and VEGFR2 activation during the EC differentiation process. QKI-5 was induced during EC differentiation, resulting in stabilisation of STAT3 expression and modulation of VEGFR2 transcriptional activation as well as VEGF secretion through direct binding to the 3’ UTR of STAT3. Importantly, iPS-ECs overexpressing QKI-5 significantly improved angiogenesis and neovascularization and blood flow recovery in experimental hind limb ischemia. Notably, human iPS cells overexpressing QKI-5, induced angiogenesis on Matrigel plug assays in vivo only seven days after subcutaneous injection in SCID mice. These results highlight a clear functional benefit of QKI-5 in neovascularization, blood flow recovery and angiogenesis. They, thus, provide support to the growing consensus that elucidation of the molecular mechanisms underlying EC differentiation will ultimately advance stem cell regenerative therapy and eventually make the treatment of vascular diseases such as diabetes a reality.

Biography:

Francesco Neri obtained PhD in Biotechnology in Siena (Italy) working on embryonic stem cells epigenetics. Then he worked as Postdoc between Human Genetics Foundation (HuGeF) in Torino (Italy) and the Radboud University Medical Centre, Nijmegen (Netherlands) where specialized his research on the DNA methylation specific epigenetic mark on stem cell, differentiation and colon cancer. During this period, he characterized new mechanisms of gene transcriptional regulation, developed new genome-wide methods and identified the role of the intragenic methylation. Since 2016, he is the Group Leader of the “Epigenetics research group” at the Fritz Lipmann Leibniz Institute on Aging (FLI) in Jena (Germany). His current research is supported by the Sofja Kovalevskaja starting grant of the von Humboldt foundation, and it is focused on the aging-dependent epigenetic aberrations occurring in adult stem cells promoting colon cancer.

Abstract:

DNA methylation is a heritable epigenetic modification required for embryonic development, which causes transcriptional repression when established on gene promoters. Recent studies have reported that Dnmt3b binds preferentially to the gene bodies by interacting with the histone modification H3K36me3. While the molecular and biological functions of intragenic DNA methylation are still unknown, the deregulation of this epigenetic feature has been associated with several diseases. Here, we show that the Dnmt3b-dependent intragenic DNA methylation protects the gene body from RNA Polymerase II (RNA Pol II) spurious entry and cryptic transcription initiations. Using different genome-wide approaches, we demonstrate that loss of Dnmt3b leads to an increase of the RNA Pol II engagement within gene bodies resulting in the onset of spurious intragenic transcription initiations. Finally, we demonstrate that inhibition of RNA Pol II spurious entry depends on the enzymatic activity of the Dnmt3b recruited by H3K36me3. Our results elucidate the functional role of the Dnmt3b-dependent intragenic DNA methylation, and the existence of a RNA Pol II-triggered epigenetic crosstalk involving SetD2, H3K36me3, Dnmt3b and DNA methylation, to ensure gene transcription initiation fidelity. This security feedback is probably lost during cancer development, where a global intragenic hypomethylation frequently occur, which increases transcription instability and promotes (stochastic) tumor cell heterogeneity.

Biography:

Raphael Gorodetsky is the head of the Laboratory of Biotechnology and Radiobiology at the Sharett Institute of Oncology, Hadassah - Hebrew University Medical Center in Jerusalem, where he is employed as a faculty member since 1989 (affiliated to the Hebrew University Medical School in Jerusalem). He received his M.Sc. and PH.D from the Hebrew University in 1985 and had his Postdoc at UCLA Medical Center (1985-1988). Among other subjects he was involved in cancer research and radiobiology studies, as well as in projects in regenerative medicine. His earlier studies were associated with trace elements physiology in health and disease. Later directions of his research focused mainly on the invention of fibrin based biomaterials used for tissue regeneration and cell therapies and on different aspects of cancer research and radiation biology. Among the new ventures he co-founded was Hapto Biotech in 2000, where he served as the chief scientist (later merged with Ortec to form Forticel International, NY). In this area he designed fibrin based matrices for tissue regeneration, specifically of bone and cartilage. In recent years he was deeply involved in the development of placental stromal cell based treatment for regenerative and tumour control purposes, including the mitigation of radiation effects. These findings were applied clinically by Pluristem Therapeutics. In parallel he is involved in the development of new anticancer immunotherapies. Besides authoring more than 100 peer reviewed scientific publications and chapters, he edited and authored the book “Stem cells and Tissue Repair” in 2010 (by the Royal Society of Chemistry RSC, Cambridge, UK). He served as secretary of Israel Stem Cells Society (2014-2017) and has been a member in a number of related national and international societies.https://stemcell-regenerativemedicine.conferenceseries.com

Abstract:

We developed a technique for direct isolation of human placenta stromal cells from tissues of fetal origin (fPSC) by a unique process which allows the direct migration of the desired cells from tissue fragments to culture dishes. These selected placental stromal cell population was found to be highly potent indirect enhancers the regeneration of failing bone marrow and mitigation of acute radiation syndrome (ARS) following total body irradiation. The study was based on our previous record with similar cells of mixed maternal and fetal origin produced by a corporate in bioreactors to fit for clinical applications. The advantage of IM treatment by fPSC was that these stromal cells were found to be more immunocompetent and could reside longer in the injected muscle with no apparent adverse effects that were described following IV delivery of MSC of different origins, where most of the injected cells were trapped in the lungs. The cell treatments induced rapid indirect therapeutic effects. In studies on mitigation of radiation effects these cells enabled to the fast repopulation of the bone marrow lineages with subsequent regeneration of the peripheral blood cells. This saved the animals from the lethal effects of ARS with dramatic significant raise in ~8Gy irradiated mice from less than a third to almost 100%, with fast recovery of the bone marrow and peripheral blood cells. Cytokines analyses showed that the injected xenogeneic cells respond to the stress of the heavily irradiated mice by secretion of a wide range of related pro-regenerative cytokines. The IM based fPSC treatments have also been investigated as cell therapy for treating bone marrow failure due to different other causes. Other indications tested for optional PSC treatment include regeneration of the salivary glands after heavy dose head and neck irradiation, regeneration of tissues affected by autoimmune diseases. These included inflammatory bowel disease (IBD) and autoimmune inflammatory processes in the brain, such as multiple sclerosis. Further detailed studies are performed to better understand the indirect mechanism of action of the PSC by stress induced activation of relevant family of genes in these cells.

Biography:

Yuval Rinkevich is the young Principle Investigator with supervising and mentoring position at the Institute for Lung Biology and Disease, Comprehensive Pneumology Center (CPC), Helmholtz Zentrum, Munich, Germany. He obtained a PhD degree in Biology from Technion of Israel Institute of Technology, Haifa in 2008. From 2008 until 2014, he was postdoctoral fellow of Prof. Irving L. Weissman, Stanford Institute for Stem Cell Biology and Regenerative Medicine at Stanford University, USA. In 2014 he became Basic Life Science Research Associate at Stanford University. He was on the Team of inventors of two Patents regarding ‘Isolation and Characterization of Progenitor Cells from Mesothelium and Methods’ and ‘Compositions for the Prevention and Treatment of Surgical adhedions’. His lab is exploring the stem cells, embryonic lineages and mechanisms by which tissues/organs regenerate following injury, at multiple levels of biological organization. His research is currently funded by grants from the Else-Kröner-Fresenius Stiftung (EKFS), Human Frontier Science Program Organisation (HFSPO), German Research Foundation (DFG) and the Fritz Thyssen Stiftung (FTS). He is member of several Scientific Societies and publishes in peer-reviewed journals, gives lectures at numerous, distinguished Conferences and Institutes as COST, GRC, ICB, TUM, EMBO, TERMIS, BMF and ATS and was recently elected to the ETRS Board.

Abstract:

All mammals and humans undergo metamorphosis in response to injury, from Regeneration To Scarring (RTS). Here we follow two functionally diverged fibroblastic lineages (ENFs & EPFs) and document their lineage successions during backskin development that coincides with RTS. We show that ENFs are dermal sculptures that develop and regenerate native architectures during fetal life, and that their lineage decline over time imposes a dermal tissue absent of such events. We show that EPFs are scar producers even at fetal stages, wherein their numbers are below a threshold needed to generate macroscopic scars, but that their dynamics predicts scar emergence. We show that clonal advantages to EPFs rather than programed cell lineage death, most likely are primary succession mechanisms, and that RTS can be partly circumvented by transplantations of fetal ENFs or decellularised fetal dermis. Our findings provide a mechanism for regenerative decline in mammals, carry clinical implications by suggesting that human dermal regeneration could be reached by coxing or transplanting ENFs alone, and provide a model for comparative regeneration studies between taxon groups.

Biography:

Yibing Li the Founder and President of Midwest Joint Pain Institute and Centre for Pain Management & Rehab, is an interventional Physiatrist. She is American board certified in physical medicine and rehabilitation, pain medicine and small fiber nerve conduction studies. She graduated from Shenyang Medical College in China and completed residency at BJH/Washington University School of Medicine at St. Louis. She has over 12 years of clinical experience in spine and joint pain management. She has special interest and extensive training in regenerative medicine and is one of frontiers in performing prolotherapy, PRP, amniotic allograft tissue and bone marrow stem cell therapies in the United States.

Abstract:

Overview: To monitor the clinical outcomes from comprehensive regenerative therapies with prolotherapy, platelet rich plasma (PRP) and human amniotic allograft matrix injections to treat patients with chronic joint pain and all degrees of osteoarthritis in an outpatient clinic setting.

Introduction: Ligament laxity, tendinopathy, cartilage degeneration and previous joint injuries or surgeries are the major root causes of persistent joint pain and osteoarthritis. Dextrose prolotherapy, PRP and amniotic allograft injection into joints and surrounding weakened ligaments and tendons have been recommended to reduce chronic joint pain, improve functional status, and correct underlying causes of osteoarthritis. The current report is to assess the clinical outcomes of these comprehensive therapies among patients who had persistent joint pain and all degrees of degenerative joint disease/OA, ligament laxity or tendinopathy from imaging studies and failed with most of other conservative treatments.

Methods:  From September 2015 to August 2017, 118 patients  who had chronic joint pain and various degrees of osteoarthritis were treated with a series of injection of prolotherapy, PRP and human amniotic allograft tissue matrix to different parts of the body (N=94, 11, 9, and 4 to knees, hips, shoulders, and ankle/foot joints, respectively) under ultrasound needle guidance. The age range of the patient population is from 38-99 years, 62 males and 56 females. 31.4% of the patients had mild to moderate degenerative joint disease (DJD) and 64.4% had severe DJD. 19.6% of the patients had history of joint surgeries. Outcomes are the difference or % of change in VAS pain scale, upper extremity DASH and lower extremity 80-point functional scales, measures obtained from physical exams, X-ray and musculoskeletal ultrasound between pre- and 2-3 months, 4-6 months, 6 months and above post-treatments.

Results: After 2-6 months post-initial treatments, on VAS pain scales, 13.5% of the patients reported 0/10 or no pain and overall 74.6% reported at least 25%-50% pain reduction. 76.2% reported improvements in LEFS scale. Almost all patients showed varied degrees of improvements in both from soft tissue regenerations on ultrasound comparison and the physical exams between pre- and post- treatments. On the X-ray comparison, 66.7% showed improvement while 33.5% showed no change. None had experienced any adverse reactions from these injections.

Conclusion: Comprehensive regenerative therapies are safe, effective, non–surgical treatment options for patients with persistent joint pain and functional limitations from soft tissue degeneration/tears and osteoarthritis. Further treatments are recommended to some patients and continuous clinical studies are warranted to evaluate the long term benefits.

Biography:

Yibing Li the Founder and President of Midwest Joint Pain Institute and Centre for Pain Management & Rehab, is an interventional Physiatrist. She is American board certified in physical medicine and rehabilitation, pain medicine and small fiber nerve conduction studies. She graduated from Shenyang Medical College in China and completed residency at BJH/Washington University School of Medicine at St. Louis. She has over 12 years of clinical experience in spine and joint pain management. She has special interest and extensive training in regenerative medicine and is one of frontiers in performing prolotherapy, PRP, amniotic allograft tissue and bone marrow stem cell therapies in the United States.

Abstract:

Overview: To evaluate the clinical outcomes of comprehensive regenerative therapies with mesenchymal stem cells (MSCs) from bone marrow, platelet rich plasma (PRP) and dextrose prolo-solution injections to treat patients with persistent joint pain and severe osteoarthritis at an outpatient regenerative medicine clinic.

Introduction: Ligament laxity and cartilage degeneration are known to be root causes of osteoarthritis. Comprehensive regenerative therapies with the combination of dextrose prolotherapy, PRP and autologous stem cell injections into the pathologic joint and surrounding weakened tendon and ligaments are effective in improving pain and overall function in all degrees of osteoarthritis. In this study from an outpatient clinic, MSCs from bone marrow aspirate with PRP and dextrose prolotherapy were used for 19 patients (14 male, 5 female, age from 30-70 years old) from Sept. 2016 to August 2017. Among them, 18 patients had severe degree of degenerative joint disease/osteoarthritis; one had severe shoulder pain from rotator cuff tendon tears. All of them failed with traditional treatments and then selected for alternative treatments to surgeries. Treatments were made with one time PRPs and MSC bone marrow stem cells injections to 20 knees, 1 shoulder, 1 ankle and 2 hip joints, respectively under imaging needle guidance. Outcomes included the changes in VAS pain scale, upper or lower extremity functional scale, X-rays and measurements from musculoskeletal ultrasound pre- and post-treatments.

Results:  After 2-6 months post initial treatments, data was collected in 18/19 patients. There were an average of 58.2% pain reduction at VAS pain scale and 46.7 % improvement in functional scales. X-ray improvements seen in 2 out of 3 patients. Over 90% patients showed varies degree of new soft tissue growths from musculoskeletal ultrasound exam comparisons. No adverse reactions or complications were observed in all patients.

Discussion and Conclusions: Comprehensive regenerative therapies, as above, are new cutting-edge treatment compared to the alternate invasive orthopaedic surgeries after failure of other traditional standard-of-care treatments. Bone marrow stem cell aspiration and subsequent joint injections with added PRP and dextrose prolotherapy are safe, cost-effective and relatively low-risk for patients with persistent joint pain and arthritis caused by torn tendons, ligaments ligament laxity and degenerative cartilages. For moderate to severe degree of joint pain and arthritis, we would recommend more rounds of regenerative therapies to improve the results. Further clinical studies are warranted to evaluate the long-term benefits.

Biography:

Jeffrey Xue is an avid learner and diligent pre-med student. He currently attends Knox College in IL, USA with major in Chemistry and minor in Chinese Study. He works at Midwest Joint Pain Institute during his summer time as a Research Assistant. He has strong interest in researching and studying stem cell therapy and regenerative medicine. He serves as a Student Athletic Advisory Committee Member at Knox and was the Founder and President of the science club at Lake Forest Academy. He has earned over 450 hours volunteer experience in the healthcare field.

Abstract:

Case Description: A 50-year-old male complained of right shoulder pain, popping and clicking with shoulder movements for one month after boxing. He couldn’t raise his right arm above his shoulder. He had difficulty sleeping at night because of the shoulder pain. He had previous right shoulder pain from rotate calf tendons and labrum tear, the pain was resolved from a series of prolotherapy and PRP injection over a year ago after failure of rest, NSAIDs, PT, etc. On the physical exam, he had reduced right shoulder range of motion to 50% normal. He had focal tenderness anterior right shoulder with (+) Hawkin’s test and (-) drop arm test. MSK ultrasound exam reviewed two large incomplete tears in the right subscapularis tendon, measured at 0.33 cm x 0.31 cm and 0.54 cm x 0.31 cm, respectively. He also had chronic and diffuse rotator cuff tendinopathy. Due to the recurrent injury and tendon tears, he decided to pursue bone marrow stem cell therapy. He received 14 cc of mesenchymal stem cells and growth factors injection from bone marrow aspiration to the torn subscapularis and other rotator cuff tendons with sterile techniques and imaging guidance at an outpatient regenerative medicine and PM&R clinic. The whole procedure lasts a few hours without IV sedation and he was discharged home without any complications.

Result: On the one month follow-up visit, he reported his right shoulder pain markedly reduced, he can sleep well at night. He can raise his right arm over the shoulder without pain. His shoulder ROM is nearly 90% normal range. On the repeated ultrasound exam, his previous torn subscapularis tendon had totally healed. In three months after the stem cell therapy, he was ready to go back to boxing.

Discussion: The adult bone marrow is a rich source of mesenchymal stem cells and growth factors. The bone marrow stem cell injection therapy allows our own body to restore, repair and regenerate damaged tissues and organs. Many research studies show that MSCs are primitive cells with capacities to self-replicate, fight apoptosis, reduce inflammation and differentiate into multiple tissues, including bone, muscles, tendons, ligaments, cartilage and fat. Due to the poor healing process from injuries or aging changes to the avascular structures, such as tendon, ligaments, cartilage, meniscus, labrums, etc., the patients usually suffer from the chronic joint pain and disability. Traditional treatments with NSAIDS, pain medications and arthroscopic surgeries are usually either not effective, lack of long term success or carrying significate risks and requiring prolonged healing time. The regenerative therapies with direct injections with MSCs and growth factors injection to those damaged tissues have clinically proven to be very simple procedures and highly effective treatments by targeting to resolve the root of chronic painful conditions with no downtime and very low risk or complications.

Conclusion: Bone marrow stem cell therapy is potentially very effective and alternative treatment to shoulder pain from incomplete rotator cuff tears and tendinopathy with very minimum risks, complications and side effects compared to current most traditional treatment options.

Biography:

Monika Glemžaitė is a second year PhD student in Biochemistry at Vilnius University. Her research is focused on epigenetic mechanisms distinguishing amniotic fluid-derived stem cells functioning. She is a co-author in the international patent application related to CRISPR/Cas9 and has been awarded a Young Inventor Medal by World Intellectual Property Organization (WIPO) for CRISPR/Cas research. She has two publications as a first author and has presented her work in six international scientific conferences. She is also actively spreading science among students and public in Lithuania.

Abstract:

Human amniotic fluid-derived mesenchymal stem cells (AF-MSCs) are fetal mesenchymal stem cells having multilineage differentiation potential and found in amniotic fluid. The aim of the present study is to assess the potency of AF-MSCs to differentiate into cardiomyogenic lineage in vitro using DNMT inhibitors, Decitabine, Zebularine, RG108 and p53 inhibitor Pifithrin-α and determine epigenetic changes during this differentiation. Cells were obtained from amniocentesis samples from second trimester women who needed prenatal diagnostics (protocols approved by the Ethics Committee of Biomedical Research of Vilnius District, No 158200-123-428-122). Isolated AF-MSCs were characterized by cell surface markers (CD44, CD90, CD105 positive and CD34 negative) and stem cells pluripotency genes (OCT4, SOX2, NANOG, REX1). Cardiomyogenic differentiation was confirmed by cell staining as well as by expression of cardio myogenesis-related genes NKX2-5, TNNT2, MYH6 and DES using RT-qPCR. MTT assay showed that all differentiation inducers and especially Pifithrin-α suppressed cell proliferation at the beginning of differentiation. Western blot analysis of epigenetic changes revealed the reduction in levels of Polycomb repressive complex 2 (PRC2) proteins EZH2 and SUZ12 and chromatin remodeling enzymes DNMT1, HDAC1/2 and HP1α after AF-MSCs induction with all agents. Also, alterations in marks both keeping active (H3K4me3, H3K9Ac and H4hyperAc) and repressed (H3K27me3 and H3K9me3) state of chromatin were observed. Our results demonstrate various changes in genetic and epigenetic profiles of AF-MSCs differentiated into cardiomyocytes progenitors resulting in global chromatin remodeling. Therefore AF-MSCs can be proposed as a potential alternative of stem cells for regenerative medicine.

Biography:

Collin Looi graduated from International Medical University in 2008. He is currently enrolled in a Master in Orthopaedic programme in Universiti Putra Malaysia. He has two published papers and is a keen researcher, with interest in Stem cells and non-unions of bone.

Abstract:

Granulocyte-Colony Stimulating Factor (G-CSF) is a native hematopoietic cytokine produced by fibroblasts, monocytes and endothelial cells, which primarily regulate neutrophil production and upregulates mesenchymal stem cell proliferation within the bone marrow and peripheral circulation. These mesenchymal stem cells have been described as key components involved in bone fracture healing. We hypothesized that by increasing the concentration of these mesenchymal stem cells by systemic administration of G-CSF, the rate of fracture healing could be expedited, potentially opening doors for further clinical applications. A caprine based animal study was done to assess the effect of systemically administered G-CSF on the rate of bone fracture healing involving 2 groups (control group and G-CSF group) with 5 animals in each group. Rate of fracture healing was assessed via radiographic methods
using a validated score measuring the parameters of bridging and union. Results revealed a median time to bridging of 7 weeks in the control group and 4 weeks in the G-CSF group (interquartile range of 0.5 weeks in both groups), while a median time to union of 13 weeks in the control group and 8 weeks in the G-CSF group (interquartile range of 1.5 weeks in both groups). A Mann-Whitney U test demonstrated a significant difference between the control group and G-CSF group in relation to bridging (p-value 0.008) and
union (p-value 0.008) times. This confirmed our research hypothesis that systemic administration of G-CSF produces an optimized fracture-healing rate.