Day 14 :
- Stem Cell Therapy
Chair
Mariusz Grudniak
Professor
Session Introduction
Sholeh Etehad Asnaf
Student at Azad University,North Tehran Branch Department of biology,Iran
Title: Evaluation of gene expression of NLRP3 and NLRC4 as Inflammasome receptors in peripheral blood of SM injured patient by Real-Time
Time : 1:30-2:15
Biography:
Abstract:
Mustard gas is one of the most widely used chemical warfare agents used during the Iraqi wars against Iranian warriors. Some mechanisms of chronic pulmonary damage caused by mustard gas such as oxidative stress, protease-antiprotease imbalance and inflammation have been studied. In the context of inflammation, the role of inflammacom and the genes involved in it, especially the receptors of this inflammatory complex, is important. Therefore, the present study evaluated the expression of NLRP3 and NLRC4 genes as inflammatory receptors in peripheral blood of veterans of pulmonary chemo using Real- Time PCR. The present study was conducted as a cross-sectional analytical study on 15 chemical warfare victims exposed to mustard gas, 15 COPD patients and 15 healthy individuals as controls that were referred to the lung clinic of Baqiyatallah Hospital (AS). After extracting RNA from the blood sample and synthesizing cDNA, expression of the two NLRP3 and NLRC4 genes was evaluated using Real Time PCR. Finally, the data were analyzed by SPSS version 20 The two NLRP3 and NLRC4 genes did not change significantly in veterans and people with COPD compared to healthy subjects. Although the expression of these two genes was higher in COPD patients than veterans (0.17 to 0.05 and 0.15 to 0.04, respectively), there was no statistically significant relationship. According to the results of the present study, it has been found that two NLRP3 and NLRC4 genes have the potential to be involved in inflammation and chronic obstructive pulmonary embolism. As a result, it is hoped that with the full knowledge of the process of chronic damage to the lungs caused by mustard gas, one can use it to create a pattern of diagnosis and prevent its further complications
Dr. Khalid Ahmed Al-Anazi
Consultant and Chairman, Department of Adult Hematology and Hematopoietic Stem Cell Transplantation
Title: Cure of Insulin-Dependent Diabetes Mellitus by an Autologous Hematopoietic Stem Cell Transplantation Performed to Control Multiple Myeloma in a Patient with Chronic Renal Failure on Regular Hemodialysis
Time : 2:15-3:00
Biography:
Abstract:
Umang G. Thakkar
Department of Stem Cell Therapy and Regenerative Medicine
Title: Co‑infusion of Autologous Adipose Tissue Derived Neuronal Differentiated Mesenchymal Stem Cells and Bone Marrow Derived Hematopoietic Stem Cells, a Viable Therapy for Post‑traumatic Brachial Plexus Injury: A Case Report
Biography:
Abstract:
Stem cell therapy is emerging as a viable approach in regenerative medicine. A 31-year-old male with brachial plexus injury had complete sensory–motor loss since 16 years with right pseudo‑meningocele at C5–D1 levels and extra‑spinal extension up to C7–D1, with avulsion on magnetic resonance imaging and irreversible damage. We generated adipose tissue derived neuronal differentiated mesenchymal stem cells (N-AD-MSC) and bone marrow derived hematopoietic stem cells (HSC-BM). Neuronal stem cells expressed β-3 tubulin and glial fibrillary acid protein which was confirmed on immunofluorescence. On day 14, 2.8 ml stem cell inoculum was infused under local anesthesia in right brachial plexus sheath by brachial block technique under ultrasonography guidance with a 1.5-inch-long 23 gauge needle. Nucleated cell count was 2 × 104/µl, CD34+ was 0.06%, and CD45-/90+ and CD45-/73+ were 41.63% and 20.36%, respectively. No untoward effects were noted. He has sustained recovery with re-innervation over a follow-up of 4 years documented on electromyography-nerve conduction velocity study.
- Regenerative Medicine
Chair
Raghvendra V Tey
Professor
Session Introduction
Xianmin Zeng
Buck Institute, Novato, CA, USA
Title: Making multiple therapeutic cell products from a cGMP-compliant iPSC line
Biography:
Dr. Xianmin Zeng received her PhD in Molecular Biology from the Technical University of Denmark in 2000 and had her postdoctoral training at the NIH. She joined the faculty of the Buck Institute Research on Aging in 2005 where she builds the Institute’s Stem Cell Program, and is current Professor at the Buck Institute. Dr. Zeng is a recipient of several major funding including a translational grant to develop clinically grade dopaminergic neurons from pluripotent stem cells for Parkinson’s disease from California Institute for Regenerative Medicine (https://www.cirm.ca.gov/our-progress/people/xianmin-zeng), and an iPSC-based toxicity screen grant from the NIH. She is also the Founder and Chief Executive Officer of XCell Science Inc, a biotech company dedicated to providing reagents and services in neural space, and NxCell Inc, an iPSC-based therapy company.
Abstract:
We have developed several current Good Manufacture Practice (cGMP)-compliant induced pluripotent stem cell (iPSC) lines for clinical applications. Here we show that multiple cellular products currently being considered for therapy can be generated from a single master cell bank of a clinically compliant iPSC line. Here we tested differentiation into therapeutic relevant cell types of the three germ layers by standardized protocols using a stock prepared from the cGMP-compliant working cell bank. Cells we generated include 1) neural stem cells, dopaminergic neurons and astrocytes, 2) retinal cells (RPE and photoreceptors) and 3) hepatocyte, endothelial and mesenchymal cells. To confirm that these protocols can also be used for other iPSC lines we tested the reproducibility of our methodology with a second clinically compliant line. Our results confirmed that the same protocols could be used with minimal modifications with multiple qualified lines. In addition, we introduced a constitutively expressed GFP cassette in Chr13 safe harbor site and observed no significant difference in growth and differentiation between the engineered line and the control line. We believe that our demonstration that multiple products can be made from the same WCB, and that the same protocols can be used with multiple lines offers a path to a cost effective strategy for developing cellular products from iPSC lines.
- Stem cells | Genetics and stem cell biology | Cancer stem cells | Tissue engineering | Stem cell therapy | Stem cell transplantation | Stem cell niche | Regenerative Medicine | Adult stem cells | Embryonic stem cells
Session Introduction
Xianmin Zeng
Buck Institute, Novato, CA, USA
Title: Making multiple therapeutic cell products from a cGMP-compliant iPSC lin
Time : 1:30-2:00
Biography:
Dr. Xianmin Zeng received her PhD in Molecular Biology from the Technical University of Denmark in 2000 and had her postdoctoral training at the NIH. She joined the faculty of the Buck Institute Research on Aging in 2005 where she builds the Institute’s Stem Cell Program, and is current Professor at the Buck Institute. Dr. Zeng is a recipient of several major funding including a translational grant to develop clinically grade dopaminergic neurons from pluripotent stem cells for Parkinson’s disease from California Institute for Regenerative Medicine (https://www.cirm.ca.gov/our-progress/people/xianmin-zeng), and an iPSC-based toxicity screen grant from the NIH. She is also the Founder and Chief Executive Officer of XCell Science Inc, a biotech company dedicated to providing reagents and services in neural space, and NxCell Inc, an iPSC-based therapy company.
Abstract:
We have developed several current Good Manufacture Practice (cGMP)-compliant induced pluripotent stem cell (iPSC) lines for clinical applications. Here we show that multiple cellular products currently being considered for therapy can be generated from a single master cell bank of a clinically compliant iPSC line. Here we tested differentiation into therapeutic relevant cell types of the three germ layers by standardized protocols using a stock prepared from the cGMP-compliant working cell bank. Cells we generated include 1) neural stem cells, dopaminergic neurons and astrocytes, 2) retinal cells (RPE and photoreceptors) and 3) hepatocyte, endothelial and mesenchymal cells. To confirm that these protocols can also be used for other iPSC lines we tested the reproducibility of our methodology with a second clinically compliant line. Our results confirmed that the same protocols could be used with minimal modifications with multiple qualified lines. In addition, we introduced a constitutively expressed GFP cassette in Chr13 safe harbor site and observed no significant difference in growth and differentiation between the engineered line and the control line. We believe that our demonstration that multiple products can be made from the same WCB, and that the same protocols can be used with multiple lines offers a path to a cost effective strategy for developing cellular products from iPSC lines.
Chandravanshi Bhawna
School of Regenerative medicine, GKVK Post, Bellary Road, Yelahanka, Bangalore
Title: Stem cell therapy facilitates tolerance in renal transplantation
Time : 2:00-2:30
Biography:
Bhawna Chandravanshi during the tenure of her project mastered several basic as well as specialized cell culture techniques such as maintenance of normal and cancer cell lines, isolation of islets from murine pancreas, isolation and cultivation of stem cells from Human umbilical cord, amnion, placenta etc. She explored the possibility of in vitro generation of islets from the mesenchymal stem cells (MSCs) derived from discarded tissues of human origin. She succeeded in obtaining large number of functional islets from these MSCs and could demonstrate restoration of normoglycemia upon transplantation of these islets in STZ diabetic mice. To her credit she has 5 first author publications, 5 second author publications with 2 book chapters. She has already visited Japan twice for oral and poster presentation and has bagged best oral presentation award.
Abstract:
The auto-immune destruction of pancreatic β cells leads to occurrence of Type 1 diabetes. Two main therapeutic approaches for the treatment of T1D are islet transplantation and exogenous insulin injection. However both the approaches have not attained significant success. One of the main reasons for the decline in the success of islet transplantation is the shortage of donor pancreas and their poor survival upon transplantation. Hence in the present study we generated ample number of islet like cell aggregates ( ICAs) from Wharton’s jelly mesenchymal stem cells (WJ-MSC) as published earlier and stored them at ultra low temperature (-196ËšC) employing a cocktail of small molecules (Eicosapentaenoic acid+Docosahexaenoicacid+Metformin)) in addition to 5% DMSO. The ICAs were revived after 30days, cultured for 24h and then were assessed for their viability, oxidative status and functionality. The viability was determined by MTT and FDA/PI assay and it was found that the combinations of small molecules maintained the viability by 4fold when compared to controls Further the small molecules guarded the islets against oxidative stress and hence significantly reduced total ROS (p<0.05) (55% reduction), superoxide ions (p<0.05) (65% reduction) and nitric oxide (p<0.01) (70%) free radicals. The expression of HIF1α was also enhanced in the presence of small molecules. There was significant down-regulation of the stress markers like CHOP, NOS2a and IL1β, apoptotic markers like p53 and Caspase 3.Further this was confirmed by western blot analysis where we found down-regulation of Caspase 3 and phosphor p38 and up-regulation of anti-apoptotic marker, BCl2. Finally the ICAs also responded to glucose and showed significant enhancement in insulin secretion in presence of high glucose (5 fold). Hence our data demonstrates for the first time that a cocktail of small molecules with only 5% DMSO protect islets against cryo injury thus opening new dimension for islet banking prior to transplantation.
Sholeh Etehad Asnaf
Azad University,North Tehran Branch Department of biology,Iran
Title: Evaluation of gene expression of NLRP3 and NLRC4 as Inflammasome receptors in peripheral blood of SM injured patient by Real-Time
Time : 2:30-3:00
Biography:
She worked in laboratory and Baghiyatollah hospital on chemical warfare victimsand and successfully defend her thesis In the title of Evaluation of gene expression of NLRP3 and NLRC4 as Inflammasome receptors in peripheral blood of SM injured patient by Real-Time She is highly experienced in different techniques including PCR, Electrophoresis, cell culture and western blot.
Abstract:
Mustard gas is one of the most widely used chemical warfare agents used during the Iraqi wars against Iranian warriors. Some mechanisms of chronic pulmonary damage caused by mustard gas such as oxidative stress, protease-antiprotease imbalance and inflammation have been studied. In the context of inflammation, the role of inflammacom and the genes involved in it, especially the receptors of this inflammatory complex, is important. Therefore, the present study evaluated the expression of NLRP3 and NLRC4 genes as inflammatory receptors in peripheral blood of veterans of pulmonary chemo using Real- Time PCR. The present study was conducted as a cross-sectional analytical study on 15 chemical warfare victims exposed to mustard gas, 15 COPD patients and 15 healthy individuals as controls that were referred to the lung clinic of Baqiyatallah Hospital (AS). After extracting RNA from the blood sample and synthesizing cDNA, expression of the two NLRP3 and NLRC4 genes was evaluated using Real Time PCR. Finally, the data were analyzed by SPSS version 20 The two NLRP3 and NLRC4 genes did not change significantly in veterans and people with COPD compared to healthy subjects. Although the expression of these two genes was higher in COPD patients than veterans (0.17 to 0.05 and 0.15 to 0.04, respectively), there was no statistically significant relationship. According to the results of the present study, it has been found that two NLRP3 and NLRC4 genes have the potential to be involved in inflammation and chronic obstructive pulmonary embolism. As a result, it is hoped that with the full knowledge of the process of chronic damage to the lungs caused by mustard gas, one can use it to create a pattern of diagnosis and prevent its further complications
Umang G. Thakkar
Department of Stem Cell Therapy and Regenerative Medicine
Title: Co‑infusion of Autologous Adipose Tissue Derived Neuronal Differentiated Mesenchymal Stem Cells and Bone Marrow Derived Hematopoietic Stem Cells, a Viable Therapy for Post‑traumatic Brachial Plexus Injury: A Case Report
Time : 3:30-4:00
Biography:
Abstract:
Stem cell therapy is emerging as a viable approach in regenerative medicine. A 31-year-old male with brachial plexus injury had complete sensory–motor loss since 16 years with right pseudo‑meningocele at C5–D1 levels and extra‑spinal extension up to C7–D1, with avulsion on magnetic resonance imaging and irreversible damage. We generated adipose tissue derived neuronal differentiated mesenchymal stem cells (N-AD-MSC) and bone marrow derived hematopoietic stem cells (HSC-BM). Neuronal stem cells expressed β-3 tubulin and glial fibrillary acid protein which was confirmed on immunofluorescence. On day 14, 2.8 ml stem cell inoculum was infused under local anesthesia in right brachial plexus sheath by brachial block technique under ultrasonography guidance with a 1.5-inch-long 23 gauge needle. Nucleated cell count was 2 × 104/µl, CD34+ was 0.06%, and CD45-/90+ and CD45-/73+ were 41.63% and 20.36%, respectively. No untoward effects were noted. He has sustained recovery with re-innervation over a follow-up of 4 years documented on electromyography-nerve conduction velocity study. (Biomed J 2014;37:237-240)
Dudhal Swati
PhD scholar at Université Paris Diderot, France
Title: Selenoprotein N, mutated in SEPN1-related myopathy, restrains muscle stem cell differentiation’
Time : 4:00-4:30
Biography:
I am an aspiring researcher (Indian), currently working as a PhD scholar at Université Paris Diderot. I have completed MSc in Molecular Biology and Human Genetics from Manipal University and an undergraduate degree in Biomedical Science (BSc Hons) from University of Central Lancashire, UK. During my studies and mainly internships, I got interested in subjects dealing with the stem cells therapeutic applications. This was the genesis of my decision to enrol for PhD and training in translational research. During my clinical training, industrial training, research institutes and public university experience; I worked on various types of stem cells eg. Human Umbilical Cord Mesenchymal Stem Cells (HUCMSC’s), differentiation studies of murine fetal mesenchymal stem cells (FMSC’s) into DA neurons, murine Muscle Stem Cells (MuSC's) and several cell lines. I also learned Stereotactic surgery and behavioural tests for mice which was useful for treatment of neurodegenerative disorders like Parkinson’s disease. (Kumar A, et al, 2016)
Abstract:
Mutations of selenoprotein N (SEPN1) cause a congenital myopathy, SEPN1-related myopathy (SEPN1-RM), characterized by severe weakness and wasting of neck and trunk muscles, scoliosis and lethal respiratory failure. SEPN1-RM has been associated with oxidative stress, reduced satellite cell population and defective muscle regeneration. To investigate the underlying mechanisms, particularly a potential role of SEPN1 in regulating the balance between self-renewal and differentiation of the satellite cell pool, we used Sepn1 KO mouse primary satellite cells and C2C12 cells knocked down for Sepn1, at different stages of differentiation (quiescent cells, myoblasts and myotubes).
Using a suspension system to generate synchronized quiescence on C2C12, we found that SEPN1 absence does not prevent cell cycle exiting and re-entering but prevents normal downregulation of myogenic factors in G0 cells and leads to higher cyclin D1 levels in quiescence conditions. Microarray, qRT-PCR and protein studies showed that SEPN1 depletion leads to significant increase of MYOG and MYOD1 expression in proliferative C2C12. Moreover, primary muscle cells from the Sepn1 KO mice showed increased myoblast fusion at early myogenic differentiation.
We are currently exploring the mechanistic pathways leading to this cell phenotype. While we found no abnormalities of the AMPK-mediated pathway, our data suggest that HDAC5 could be involved in the accelerated differentiation phenotype. Other mechanistic studies are in progress.
In conclusion, lack of SEPN1 leads to incomplete quiescence and accelerated myogenic differentiation. Thus, we identify SEPN1 as a novel regulator of the muscle progenitor’s cell fate decision process, and SEPN1 depletion favors differentiation over self-renewal. These results potentially explain the depletion of the satellite cell population and the regeneration defect in SEPN1-RM models, and identify novel biomarkers useful to assess potential therapeutic interventions.
- Stem cell nanotechnology
Session Introduction
Mohammad Reza Hashemzadeh
PhD Candidate, Department of Stem Cells and Regenerative Medicine, Royesh Stem Cell Biotechnology Institute, Mashhad, Iran
Title: Stem Cells and Regenerative Medicine: The Potential Aspects in Cardiovascular System
Time : 9:30-10:00
Biography:
Mohammad Reza Hashemzadeh started his research studies on stem cells from 2005. In 2010, He founded the first biotechnology institute in the ministry of labor and social welfare of Iran, with cooperative of Khorasan Technical and Vocational Training Organization, in Mashhad called Eram biotechnology institute and now he is the head of stem cell department in that institute. Furthermore, He is the founder and director of ROYESH Stem Cell Biotechnology (RSCB) institute in Mashhad. He is interested in stem cells and regenerative medicine especially in cardiovascular diseases. His recent study is the evaluation of TLRs during cardiomyocyte differentiation in order to optimization of regenerative medicine in cardiovascular diseases. Also study of homing of adipose derived MSCs, study of gene regulation in stem cells based on miRNAs and siRNA and identification of a new stem cell line in Rabbit namely blastema stem like cells are another researches which he has done.
Abstract:
Stem cells have a key role in therapeutic field in today’s world. Regenerative medicine is a vast realm in which stem cells are recruited as source of tissue regeneration in most of organs especially in heart and cardiovascular system where cardiovascular diseases are the leading cause of mortality in recent years. The numerous types of stem cells such as embryonic stem cells and adult stem cells have been used in regenerative medicine especially in cardiovascular diseases and there are different routes of delivering cells to heart as a target tissue. Notwithstanding that using stem cells is a decent procedure, in some cases this is inefficient due to lack of targeted localization. This issue can be solved with triggering of CXCR4/SDF1 axis in these cells in order to stimulating of targeted mobilization and directing towards target tissue. Another aspect of regenerative medicine is the up-regulation and downregulation of microRNAs during stem cell differentiation and in cardiovascular diseases. On the other hand, the base of the majority of myocardial ischemia is inflammation and it sounds that the evaluation and manipulation of some immunological key genes such as toll like receptors and downstream elements in differentiation of stem cells into cardiomyocytes can be appropriate in therapeutic targets of cardiovascular diseases.
Jonathan Schwartz
President, Xanadu Stem Cell Clinic and Research Group
Title: Innovations and Breakthroughs for Enhancement of Stem Cell Procedures
Time : 10:30-11:00
Biography:
Jonathan Schwartz has over 37 years in Healthcare, Globalizing Emerging Medical Technologies, and has devoted the past eight years to Commercializing Cellular Therapies and Regenerative Medicine Technologies. He has been instrumental in introducing new technologies worldwide and is universally acknowledged as a pioneer in emerging medical technology. He has lectured extensively, edited publications and authored many articles.
Abstract:
Innovations and Breakthroughs for Enhancement of Stem Cell Procedures
The topic of this presentation will be devoted to recent developments in technology to enhance Cellular Therapies and what is currently commercially available.
Technologies include the following:
Nitric Oxide
Intraveneous Laser Blood Irradiation
Shock Therapy
Molecular Hydrogen Therapy
New Methods of Harvesting Stem Cells from
- Adipose Tissue
- Bone Marrow
- Dental Pulp
Supplements
Exosomes
Cytokines
How they can be incorporated and combined to enhance Stem Cell Procedures
- Stem cell in drug development
- Stem Cell Therapy
Session Introduction
Tomasz Oldak
Polish Stem Cell Bank, Warsaw, Poland FamiCord Group
Title: New insights into Alzheimer disease and potential mesenchymal stromal cell therapy
Biography:
Tomasz OÅ‚dak has completed his PhD at the age of 29 years from Institute of Hygiene and Epidemiology, Warsaw, Polamd. He is a director of Research and Development Department of Polish Stem Cell Bank. He has published 22 papers in reputed journals.
Abstract:
Alzheimer’s Disease is a non treatable condition affecting more and more people every year. There are many clinical trials concerning treatment of this disease, but none has finished with positive results so far. The aim of this study was to evaluate possible correlations between Alzheimer’s Disease and the alterations in cytokine/chemokine level in the serum of 20 patients in the various stage of the disease and 10 healthy volunteers as a control. The stem cell therapy is a very fast developing branch of medicine. Very interesting from the scientific point of view seem to be the co-culture of stem cells and limfocytes obtaining from patients and healthy donors. Cord tissue was the source of mesenchymal stromal cells (MSC). Investigated interactions between MSC and limfocytes was measured by obtaining supernatant after 1, 3, and 7 days of co-culture. The conditions allowed or preserve of direct cell-to-cell interaction from paracrinne effect due to physical barriers. The preliminary findings of this investigation showed interesting relations between cells resulting in cytokine levels. One of the proposed explanation of beta-amyloid plaques creation is the dysregulation of immunological system. Th2 lymphocytes cannot compensate increased activity of Th1 lymphocytes and their overproduction of proinflamatory cytokines. This situation has the negative outcomes. It is the inability to control the production of beta-amyloid that in particular is not eliminated and leads to neuronal death. Additional is microglia incorrect activity which is able in physiological conditions to establish protein homeostasis in central nervous system.