Day 1 :
- Stem Cells Biology | Stem Cell Therapy | Induced Pluripotent Stem Cells | Advancements and Application of Stem Cells | Hematopoietic and Embryonic Stem Cells
Geocord Cord Blood Bank, Georgia
Gocha Shatirishvili is Medical Director of family Cord Blood Bank Geocord, Tbilisi, Georgia and Chief Scientific Officer of Department for Cell Technologies and Therapy at Cancer Research Center, Tbilisi. He has earned his Doctoral Degree at the Medical University Lubeck (Germany). Since 2000 his research interests focussed on stem cell research and therapy. His 18 years long experience includes cord blood banking, cell therapy with autologous bone marrow stem cells (spinal cord injury, liver cirrhosis, heart failure, autism) and cord blood stem cells (ongoing study on autism), immunotherapy with lymphokine activated NK cells and DC vaccine against cancer. His research field includes hematopoietic stem cell expansion (cord blood), MSC expansion (from Wharton’s jelly and bone marrow). He is a member of Georgian National Committee on Transplantation and Committee on Bioethics at State Medical University Tbilisi. He is a member of Cord Blood Association (Government and Global Affairs Committee, Geneva, IL, USA) and Perinatal Stem Cell Society.
Autism Spectrum Disorder (ASD) is a neurodevelopmental condition with onset early in life. The stereotypical behavior, impairment of social communication and restricted activities are characteristics. Immuno dys-regulation, hypo-perfusion of brain and neuro-inflammation play role in pathogenesis. Lack of communication between the brain regions, responsible for normal activities, is the main pathophysiological characteristics. There is no cure for autism. ABA therapy is not enough efficient and new therapies are needed. Several trials demonstrated that CB transplantation for autism is safe and effective. Cord blood stem cell has positive paracrine effect on affected brain. But not all cord blood units contain recommended cell doses for intravenous treatment. Intrathecal injection, accepted route for neurodegenerative diseases cell therapy, could solve this problem. This is safe route, no neural tissue damage occurs, the circumvention of the blood-brain-barrier finds place. All Injected cells are transported by CSF to the affected parts of brain, while most of intravenously transplanted cells are caught in lungs, spleen and liver, only tiny number of cells migrates through blood-brain barrier. The number of cells, migrated to the brain, is important for successful treatment. Mardaleishvili Medical Center, Tbilisi startet I phase study. Children at age of 4-12 years are injected intrathecally with autologous cord blood three times with 6 months intervals. No minimal cell dose was established. Cell dose 54x106 nucleated cells per injection were efficient. 3 patients were treated yet. Effectiveness was confirmed by VABS-II and ADOS-2. No significant side effects were observed. After two transplantations children conditions were improved steadily and significantly. Behavioral improvement, less aggressiveness, no emotional outbursts and improvement of vocabulary (from zero up to thousand words) were reported. Abilities to speak, read, write, paint appeared in all three patients.
Baylor College of Medicine and Houston Methodist Hospital, USA
Dr. Kamble is Professor of Medicine in Hematology-Oncology, Cell and Gene Therapy, Baylor College of Medicine and Methodist Hospital, Houston, TX. He has served as Assistant Professor of Medicine and Associate director of Hematology-Oncology fellowship program at Oklahoma University Health Sciences Center (OUHSC). Dr. Kamble is primarily interested in hematologic malignancies including multiple myeloma and hematopoietic stem cell transplantation. He was conferred Union against cancer (ICRETT, Geneva) award in 1994 to study molecular aspect of chronic myelogenous leukemia and bone marrow transplant in Hammersmith hospital, London and Central Science and Industrial Research (CSIR) award in 1995 to study stem cell transplantation in Cornell University, New York. Dr. Kamble is board certified in Internal Medicine and Medical Oncology. He has published extensively in prestigious journals including, Blood, Biology of Bone marrow transplantation and Journal of Clinical Oncology and regularly reviews manuscripts for Blood, Biology of bone marrow transplantation and Bone marrow transplant. Dr. Kamble's clinical interests are in innovative reduced conditioning protocols, in vivo T-cell depletion and novel approaches to reduce and treat graft-versus-host disease.
Objective: Dominant cardiac involvement by primary systemic amyloidosis (AL) precludes effective AL treatment and is associated with short survival. We evaluated long term outcomes of these patients receiving Orthotopic Heart Transplantation (OHT) and Autologous Hematopoietic Stem Cell Transplantation (ASCT).
Methods: Between January 2009 and July 2018, total of 14 patients who presented with severe cardiac dysfunction as their major manifestation of AL underwent OHT. Eight of these 14 patients received ASCT. All patients had end stage heart failure and developed cardiogenic shock requiring intra-aortic balloon pump support (median 20 days, range 10-165) as a bridge to OHT.
Results: The median age at AL presentation was 54 years (42-63) in 7 females and 7 males. At median follow-up of 55 months (1-104) from OHT, 10 (71 %) patients are alive. Two patients died of post-operative complications at 1 and 7 months post OHT; 2 patient died 36 and 104 months after OHT (23 and 91 months post ASCT) of AL progression. Eight patients received ASCT at median of 13 months (13-34) after OHT. Treatment for disseminated cryptococcus delayed ASCT in one patient (#8). One patient awaits ASCT in June 2018. In the remaining 3 patients ASCT was not feasible due; to low DLCO (n=2) and prior ASCT (n=1). All 8 patients with ASCT were on tacrolimus and prednisone at the time of stem cell mobilization and hematopoietic transplant; two patients were also receiving mycophenolate mofetil and valganciclovir. We collected 4.0, 5.7, 6.1, 6.2 and 9.6 x 106 /kg CD-34+ cells in 2 days after filgrastim administration (5 ug/kg, twice daily) and plerixafor (16 mg/kg based on day- 4 CD-34+ counts) in 5 subjects. The fifth patient initially failed to mobilize but 4.3x106 /kg CD-34+ cells were subsequently obtained after stopping mycophenolate mofetil for 4 weeks. The median creatinine clearance at the time of ASCT was 42 (30-53) ml/minute. All 8 patients received a renal adjusted dose of melphalan at 140 mg/m2. Mycophenolate mofetil and valganciclovir were withheld during neutropenia until engraftment. No patients received post-transplant filgrastim. Median duration of hospitalization was 18 (15-20) days. Six patients achieved hematologic complete remission while 2 patients had a partial response following ASCT. Post ASCT reactivation of CMV was seen in 4 patients. Median survival from initial AL diagnosis is 44 (11-136) Months.
Conclusion: The strategy of OHT followed by ASCT is therefore feasible in select patients with dominant cardiac involvement and advanced heart failure.
University Hospital TÃ¼bingen, Germany
Dr. Alexander has her expertise in bone tissue engineering. The aim of her research focus is to develop an optimal stem cell source and appropriate biomaterials for future regeneration of maxillofacial bone defects. Therefore, her research group characterizes jaw periosteal cells and their in vitro osteogenic capacity and establishes optimal culture conditions and biofunctionalization strategies for biomaterials in view of future clinical performance. Related thereto, the establishment of suitable cell imaging approaches in order to assess cellular dynamics and biochemical changes of living cells represents a present focus of Alexander’s work group.
Statement of the Problem: For clinical applications of stem cell-based therapies, animal component-free culture conditions are required. In previous studies, we detected robust mineralization of Jaw Periosteal Cells (JPCs) under cultivation with clinical approved Human Platelet Lysate (hPL) compared to FCS conditions. In the present study, we performed qualitative analyses of the extracellular matrix (ECM) formed by JPCs under hPL and FCS culture conditions.
Methodology & Theoretical Orientation: JPCs were induced osteogenically under FCS and hPL culture conditions. At the end of osteogenic differentiation, biochemical composition of the formed precipitates was analyzed by Raman micro-spectroscopy and mechanical properties were assessed by Atomic Force Microscopic (AFM) measurements.
Findings: By Raman analyses, higher phosphate, lower carbonate content and higher crystallinity of hydroxyapatite minerals were detected under hPL culture conditions. Furthermore, regarding the quality of the collagen network, higher ratios of proline/hydroxyproline and higher collagen cross-linking were detected in hPL-cultured JPCs. Since hPL-supplementation leads to nearly equal production of the precursor protein proline and the mature protein hydroxyproline, these findings might indicate higher elastic properties of the collagen network in contrast to the FCS supplementation. However, cross-linking seems to be higher under hPL culture conditions. When areas of cell monolayers without precipitates were monitored by AFM, higher Young’s modulus of untreated and osteogenically induced JPCs under FCS compared to hPL culture conditions were detected. The opposite was the case for the formed precipitates: under hPL conditions, precipitates formed by JPCs showed significantly higher Young’s modulus than those formed under FCS culturing.
Conclusion & Significance: In the present study, we detected significant differences in the biochemical composition and the mechanical properties of the mineralized extracellular matrix formed by hPL and FCS-cultured JPCs. The combined technologies represent an optimal tool to assess the quality of in vitro formed bone tissue.
University of Barcelona, Spain
Dr. Michael Edel is a Group Leader and an accredited Associate Professor at the University of Barcelona, Faculty of Medicine with rolling Spanish national project grant funding. He completed his Ramon y Cajal in 2017 with I3 accreditation as distinguished Investigator. He is an expert in cancer genetics and cell pluripotency. His current research is concerned with the role of the cell cycle in attaining a pluripotent state, neural stem cell development and the development of cancer. His work has appeared in Cell (2005), Nature Biotechnology (2008) and Genes & Development (2010) and two of his publications are in the world’s top 1% most cited. He leads a group dedicated to developing new methods to make genetically stable high quality clinical grade stem cells and pluripotent stem cells to study human cardiac and neural disease with an extensive international and national collaboration.
The use of the induced pluripotent stem cell (iPSC) model is a powerful tool to study the biochemical, molecular biology and cell biology of human physiology and pathology. We have improved the model by developing a synthetic mRNA transfection method using clinical grade cell culture materials that is robust, efficient and maintains parental cell CNV. Human iPSC made with our new method and derived neural stem cells (iNSC) have significantly reduced genetic instability: (1) Lower multi-telomeric signal, (2) reduced double strand DNA breaks (DSB) measured by H2AX expression, (3) correct nuclear localization of Rad51 protein expression and (4) Whole Genome Sequencing (WGS) reveals reduced DNA structural variations implicating less co-amplifications of cancer genes. In vivo analysis demonstrates reduced teratoma growth and proliferative rate of our iPSC and iNSC. The iNSC derived from our iPSC engraft in a spinal cord injury microenvironment, have a high percentage of survival, differentiate properly to neurons, oligodendrocytes and astrocytes over time with no evidence of pathology. In conclusion, we have developed a method to generate iPSC to generate improved genetically stable iPSC and iNSC for future applications to accurately model and understand human disease.
Glycation reaction is the major cause of aging. The enhanced generation and accumulation of Advanced Glycation End Products (AGEs) from glycation reaction can cause the proteins become cross-linked, have been linked to increased risk for micro vascular and micro-vascular complications associated with diabetes mellitus, neurodegenerative diseases, inflammatory diseases, autoimmune diseases, cancer and aging. De-glycating molecule or AGE-crosslinks breaking molecule can break down crosslinking of proteins in human body that lead to pathophysiological mechanism has been recognized as a causative factor in diabetic complications, age-related diseases as nephropathy, retinopathy, elderly impaired macro and micro vascular-related diseases, Alzheimer disease and skin aging. In our study, we summarized results of de-glycating molecule or AGE-crosslinks breaking molecule application with 14 patients in anti-aging and integrative medicine clinic, having risk of metabolic syndromes as diabetes, dyslipidemia, hyperthyroid and skin aging. After regular intake of de-glycating molecule or AGE-crosslinks breaking molecule with loading dose 24,800 de-glycating unit for 2 weeks and maintenance dose 12,400 de-glycating unit continuously for 6 months. There are significant result of well controlled parameter of blood chemistry profile in diabetes, dyslipidemia and hyperthyroid as blood sugar, Hb a1c, cholesterol, triglyceride, HDL, LDL, T3 and TSH. There are good satisfaction result in skin aging as improve skin elasticity and brown spots. Moreover, there are good results of improvement in general health as good sleep, good recovery after exercise, better joints movement and improved sexual function. De-glycating molecule or AGE-crosslinks breaking molecule can be the application in anti-aging practice for longevity, particularly in patients who are risk or having metabolic syndrome.
Day 2 :
- Stem Cell Technologies | Hematopoietic and Embryonic Stem Cells | Tissue Science and Engineering | Regenerative Medicine | Cellular and Gene Therapy
Center of Regenerative Medicine in Barcelona CMR[B], Spain
Alessandra Giorgetti has completed a Bachelor in Biology (1998) and PhD in Molecular Medicine from the University of Milan, Italy. She has pursued Postdoctoral training at the GMP facility, Ospedale Maggiore Policlinico Milano and in Dr. Rafii’s group at Weill Cornell Medical College in New York from 2004 to 2008, where she acquired invaluable expertise in human hematopoietic stem cells, with a particular interest on cord blood stem cells (CBSCs).Then she joined the CMR[B] as a Research Associate at the Stem Cell Bank from 2008 to 2011. In 2012 she started her period in Inbiomed as head of the Laboratory of Hematopoiesis and Blood Disorders. In 2016 she moved to CMR[B] as a Ramón y Cajal researcher and her long-term goal is to identify developmental pathways that are involved in blood stem cells (hematopoietic stem cells, HSCs) specification during early stage of human embryo development, to direct the differentiation of human induced pluripotent stem cells (iPSCs) into clinically relevant blood stem cells.
In vertebrates, GATA2 is a master regulator of hematopoiesis, repeatedly used throughout embryo development and the adult life. Although it is well established that GATA2 is essential for the onset of mouse hematopoiesis, its role during early human hematopoietic development remains elusive. By combining time-controlled overexpression of GATA2 with genetic knockout experiments, we found that GATA2, at the mesoderm specification stage, promotes the generation of hemo-genic progenitors and their further differentiation to hematopoietic progenitor cells, while negatively regulating cardiac differentiation. Surprisingly, genome-wide transcriptional and chromatin immune-precipitation analysis showed that GATA2 bound preferentially to regulatory regions, and repressed expression, of cardiac development-related genes. In contrast, genes important for hematopoietic differentiation were up-regulated by GATA2 in a mostly indirect manner. Collectively, our data reveal a previously unsuspected role of GATA2 as a direct repressor of cardiac fates, and highlight the importance of coordinating the specification and repression of alternative cell fates.
Joel I Osorio is an innovative businessman with a distinct entrepreneurial mindset concentrated adding value on areas of Biotechnology (mRNA), Reprogramming & Regenerative Medicine for translational use in humans and a variety of clinical applications aimed for both the private and the public health sectors. He is the Founder, President and CEO of RegenerAge Clinic and RegenerAge Beauty initiatives for transnational implementations, Vice President and International Clinical Developer for Bioquark, Inc
Bioquantine® a mRNA extract from Xenopus laevis frog oocytes (purified from intra- and extra-oocyte liquid phases of electroporated oocytes), showed potential as a treatment for a wide range of conditions in animal models, including Spinal Cord Injury (SCI) and Traumatic Brain Injuries (TBI) among others. The current study observed beneficial changes with Bioquantine® administration in a patient with severe SCI. Pluripotent stem cells have therapeutic and regenerative potential in clinical situations CNS disorders. One method of reprogramming somatic cells into pluripotent stem cells is to expose them to extracts prepared from Xenopus laevis oocytes. Due to ethical reasons and legal restrictions we selected a No Option patient, deciding to include in our protocol the RestoreSensor SureScan to complete it. Based on the electrical stimulation for rehabilitation and regeneration after spinal cord injury published by Hamid and MacEwan, we designed an improved delivery method for the in-situ application of MSCs and Bioquantine® in combination with the RestoreSensor® SureScan®. To the present day the patient who suffered a complete section of spinal cord at T12-L1 shows an improvement in sensitivity, strength in striated muscle and smooth muscle connection, 14 months after the first Bioquantine® and MSCs treatment and 9 months after the placement.
REGEMAT 3D, Spain
Cartilage is a dense connective tissue with limited self-repair properties. Currently, the therapeutic use of autologous or allogenic chondrocytes makes up an alternative therapy to the pharmacological treatment. The design of a bio-printed 3D cartilage with chondrocytes and biodegradable biomaterials offers a new therapeutic alternative able of bridging the limitations of current therapies in the field. We have developed an enhanced printing processes-Injection Volume Filling (IVF) to increase the viability and survival of the cells when working with high temperature thermoplastics without the limitation of the scaffold geometry in contact with cells. We have demonstrated the viability of the printing process using chondrocytes for cartilage regeneration. This development will accelerate the clinical uptake of the technology and overcomes the current limitation when using thermoplastics as scaffolds. An alginate-based hydrogel combined with human chondrocytes (isolated from osteoarthritis patients) was formulated as bioink-A and the polylactic acid as bioink-B. The bioprinting process was carried out with the REGEMAT V1 bio-printer (Regemat 3D, Granada-Spain) through a IVF. The printing capacity of the bio-printing plus the viability and cell proliferation of bio-printed chondrociytes was evaluated after five weeks by confocal microscopy and Alamar Blue Assay (Biorad). Results showed that the IVF process does not decrease the cell viability of the chondrocytes during the printing process as the cells do not have contact with the thermoplastic at elevated temperatures. The viability and cellular proliferation of the bio-printed artificial 3D cartilage increased after 5 weeks. In conclusion, this study demonstrates the potential use of Regemat V1 for 3D bio-printing of cartilage and the viability of bio-printed chondrocytes in the scaffolds for application in regenerative medicine.
National University of Ireland Galway, Republic of Ireland
Mirella Richman is a Research Assistant in wound care with the School of Nursing and Midwifery, National University of Ireland (NUI) Galway. She has received MSc Degree in Clinical Research from NUI Galway and a BSc (Honors) in Applied Biology and Biopharmaceutical Science from Galway-Mayo IT. She is an enthusiastic researcher aimed at developing therapies for current health issues, especially by means of regenerative medicine.
Apart from being used for Hematopoietic Stem Cell (HSC) transplantation, the Mesenchymal Stromal Cell (MSC) populations in human Bone Marrow (BM) are being harnessed actively in clinical trials given their unique regenerative potential.
Several studies have been published with regards to why Healthy Volunteers (HV) donates their BM for HSC transplantation. Predominant motives include: Altruism, normative considerations, empathetic feelings for the recipient/recipient's family, weighing of the risks and benefits associated with the procedure, and social obligation.
Multiple sites are accessed to aspirate large volumes of BM (maximum 1500 ml) from donors for HSC transplantation and has been correlated with serious post-donation symptoms, which restrict donors for weeks: Excessive pain at the aspiration site, fatigue, severe post-spinal headache, fever, fainting, unexpected hospitalization and difficulty/pain while walking or sitting. A paucity of published data exists on the motivations of HV for donating their marrow for MSC basic research and the experienced side effects following the procedure.
HV that previously donated their BM (30 ml) and provided informed consented (n=48) were administered a questionnaire to identify the order of their motivations for donating their marrow for basic research and the side effects they experienced following the procedure.
The considered motivations resulted in the following descending order: To advance research for the benefit of future patients, compensation for participation, the research question was interesting, and lastly to avail of a free medical check-up. Of the 35 HV that experienced any side effect, pain, fatigue, nausea, and low blood pressure were reported - which all resolved within six days on average. No case of bleeding, fainting, vomiting, skin rash, infection or hospitalization was reported.
Donating BM for MSC basic research was primarily for future benefit of patients. The aspiration procedure can be deemed relatively safer compared to that for HSC transplantation – due to lower volumes of BM aspirated from a single collection site.
AL Fordaws Association for Women and Child & Development, Gaza Strip
The study focused on the negative effects of the infertility issue in the Palestinian society in the Gaza Strip and its impact on the psychological morale on men and women. The study aimed at the society's view in general and family members in particular on the behavior of the young man or girl. Modern medicine and alternative medicine, to treat infertility, modern medicine has customers and alternative medicine or Maismi according to customs and traditions herbal medicine, especially the treatment of "honey" also has customers. The study also focused on the negative effects of infertility in the Arab and Islamic worlds and Jaber. The study found that many cases of divorce were often caused by the encouragement of family members to the husband or other infertile or vice versa to seek divorce due to non-reproduction. One of the reasons for the divorce is also the shyness of going to the doctor and saying only, "This is God's command." There are many men who preferred to be treated. According to the herbal medicine and the reason for shyness without consulting the specialist doctor. The study concluded that the people who went to the doctor or the competent medical clinics for procreation obtained positive results through the cultivation of the tubes by the needle microscopy, insert sperm by needle and the children were happy now. The study also included Israeli measures that contributed to raising the rate of infertility in the Palestinian territories. The percentage has increased in recent years, especially in the southern West Bank, where there are areas that have been affected most because of the way the Israeli authorities bury the waste. The study also concluded that there are other environmental influences and their effect on stem cells. The study found that male infertility increased by more than 60% in men, 20% in women and 20% in unknown females. The study also showed the effect of asbestos tablets on male and female infertility.