Duke-NUS Medical School, Singapore
Human embryonic stem cells (HESC) are considered the gold standard as a cellular source for regenerative medicine. The HESCʼs are obtained from supernumerary human in vitro fertilization (IVF) embryos that cannot be used for infertility treatment. FDA and EMA require that therapeutic HESC-derived cells should be differentiated under fully human and defined conditions, the differentiation protocols should be highly reproducible, the transplanted cells should exhibit normal function in vivo and improve function of the tissue to be repaired. Also, the cells should not be tumorigenic. We developed a technique for clonal establishment and expansion of HESCs on stem cell niche laminins LN-511 or LN-521 from 8-16-cell IVF embryos under fully xeno-free and chemically defined conditions. Important from an ethical standpoint is that the IVF embryos need not be destroyed like when making cell lines from the inner cell mass of human blastocysts. By culturing the pluripotent HESCs on specific cell type specific recombinant laminins, we can differentiate the cells to various lineages such as endothelial cells, cardiomyocytes and progenitors, retinal RPEs and photoreceptor progenitors. Cardiomyocyte progenitors injected into the heart infarction region of mice for human cardiomyocyte fiber bundles resulting in improved heart function. Also, laminins specifically present in the matrix surrounding photoreceptors allow generation of progenitor cells that express typical photoreceptor markers. The differentiated cells not exhibit signs of teratoma formation. Together with a major pharmaceutical company, we are developing HESC-derived cell lines for treatment of heart injury and macular degeneration.
Karl Tryggvason, MD, PhD is Professor at Duke-NUS Medical School, Singapore and Duke University, North Carolina, as well as Senior Professor at Karolinska Institutet in Stockholm. His research concerns the molecular nature, biology and diseases of basement membranes (BM), a special compartment of the extracellular matrix. His group has cloned almost all human BM proteins and clarified genetic causes of many BM-associated diseases, such as Alport and congenital nephrotic syndromes, junctional epidermolysis bullosa and congenital muscular dystrophy, as well as studied matrix metalloproteinases, including the discovery and crystal structure of MMP-2. His group has produced most laminins as recombinant human proteins and currently the group mainly studies how different laminin isoforms influence cell growth and stem cell differentiation. Tryggvason has published over 400 research articles. He is a member of the Finnish Academy of Sciences and the Swedish Royal Academy of Sciences, and has served for 18 years as a member of the Nobel Assembly and Committee for Physiology or Medicine at the Karolinska Institute. He has received several international awards, and he is co-founder of Bio Lamina AB, Stockholm, that produces laminins for cell biology and cell therapy purposes.
Revitacell Clinic, Spain
The primary cause of death among chronic diseases worldwide is ischemic cardiovascular diseases, such as stroke and myocardial infarction. Recent evidence indicates that adult Mesenchymal stem cells therapy aimed at restoring organ function, and cardiovascular repair represent promising strategies to treat cardiovascular diseases, and have been recognized as one of the potential therapeutic agents, following several tests in animal models and clinical trials. In the process, various sources of mesenchymal stem cells have been identified which help in cardiac regeneration by either revitalizing the cardiac stem cells or revascularizing the heart. Although mesenchymal cell therapy has achieved considerable admiration and promising therapeutic strategy is the priming of therapeutic MSCs with stem cell modulators before transplantation therapeutic efficacy of MSCs In vitro or In vivo from cell priming to tissue engineering strategies, for use some challenges still remain that need to be overcome in order to establish it as a successful technique, questions going on: Which specific types of stem cells are likely to be most effective? Can heart cells divide, and if so, can we develop strategies to stimulate the growth and differentiation of the cardiac cells left in the injured heart to promote recovery of tissue mass and function?
Nobody knows at the time being what will be the best therapy for our patients. “We may need different cells for different patients and different cells for drug discovery or tissue engineering.” Which cell(s) will ultimately prove to be useful in patients is a matter of opinion.
Dr. Miguel G. Garber has over 32 years of experience in Internal Medicine and Cardiology, in addition to training, research, and development expertise in Regenerative Medicine. Over the past 12 years, he has made a significant contribution to stem cell research, specializing in the exploration and development of stem cell therapies for cardiac disorders, osteoarthritis, and neurological and autoimmune diseases. Formerly the Director of American Medical Information Group, he now serves as the Medical Director of Regenerative Medicine Madrid and the President of the Spanish Society of Regenerative Medicine and Cell Therapy (SEMERETEC). He also teaches a Masterʼs degree program in Regenerative Medicine and edits a number of scholarly journals on the subject.
Department of Medical, Chieti University, Italy
In the bone regeneration field, properties of 3D scaffold could be improved using cellular and their released products. The aim of the study was to investigate the properties of 3D printed PLA scaffolds (PLA) for bone regeneration obtained through 3D printing, evaluating the differences in terms of structural properties, in vitro and in vivo cellular responses induced by different scaffold structures. Five porous scaffold designs were fabricated from a poly-(lactide) (PLA) filament. Scaffold structural parameters were measured using scanning electron microscopy, and micro-computed tomography. Nano-topographic surface features were investigated by means of atomic force microscopy. After 112-day period, PLA were degraded and changes in weight, pH and mechanical properties. Influence of degradation on cellular activity was evaluated by means MTT assay on human periodontal ligament stem cells (hPDLSCs) in presence of degradation byproducts. Osteogenic differentiation of hPDLSCs on different scaffold designs after 21 days of culture was measured by means RT-PCR and Western Blot. After in vitro evaluation the in vivo performance were tested. In vivo study was performed using C57BL/6 mice and was designed in 5 different groups:
- Group1: PLA loaded with hPDLSCs;
- Group2: PLA loaded with Conditioned Medium (CM) derived from hPDLSCs;
- Group3: PLA loaded with Extracellular Vescicles(EVs) purified fromCM;
- Group4: PLA loaded with EVs covered with PolyEthylenImine (PEI);
- Group5: PLA, used as control.
Histological analysis were performed after 60 days of in vivo implantation and morphological evaluations revealed a high bone tissue formation and osteogenic cells commitment in group 3 and 4 when compared to other groups.
Dr. Francesca Diomede is a Research Fellow at the Department of Medical, Oral and Biotechnological Sciences, Chieti University (Italy). She received her Ph.D. in Basic and Applied Medical Sciences at University of Chieti, where she completed the postdoctoral training in stem cell biology.
REGEMAT 3D, University of Granada, Spain
Tissue regeneration (TR) is currently one of the most challenging biotechnology unsolved problems. Tissue engineering (TE) is a multidisciplinary science that aims at solving the problems of TR. TE could solve pathologies and improve the quality of life of billions of people around the world suffering from tissue damages. New advances in stem cell (SC) research for the regeneration of tissue injuries have opened a new promising research field. However, research carried out nowadays with two-dimensional (2D) cell cultures do not provide the expected results, as 2D cultures do not mimic the 3D structure of a living tissue. Some of the commonly used polymers for cartilage regeneration are Poly-lactic acid (PLA) and its derivates as Poly-L-lactic acid (PLLA), Poly(glycolic acids) (PGAs) and derivates as Poly(lactic-co-glycolic acids) (PLGAs) and Poly caprolactone (PCL). All these materials can be printed using fused deposition modelling (FDM), a process in which a heated nozzle melt a thermoplastic filament and deposit it in a surface, drawing the outline and the internal filling of every layer. All this procedures uses melting temperatures that decrease viability and cell survival. Research groups around the world are focusing their efforts in finding low temperature printing thermoplastics or restricted geometries that avoid the contact of the thermoplastic and cells at a higher temperature than the physiologically viable. This has mainly 2 problems; new biomaterials need a long procedure of clearance before they can be used in clinical used, and restrictions in geometries will limit the clinical application of 3D printing in TE.
Keywords: Bioprinting, cartilague regeneration, Poly caprolactone, scaffold
José Manuel Baena, research associate “Advanced therapies: differentiation, regeneration and cancer” IBIMER, CIBM, Universidad de Granada. Founder of BRECA Health Care, pioneer in 3D printed custom made implants for orthopedic surgery, and REGEMAT 3D, the first Spanish bioprinting company. Expert in innovation, business development and internationalization, lecturer in some business schools, he is passionate about biomedicine and technology. In his free time he is also researcher at the Biopathology and Regenerative Medicine Institute (IBIMER).