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Presenter: Alfredo, Gorio, Milano, Italy
Authors: Alfredo Gorio
Transplantation of NSCs to the injured CNS improves neuronal survival and functional outcome in short-medium term experiments are fundamental features already well demonstrated. However, longer term outcomes pose serious problems.Pluripotent embryonic stem cells have the potential to generate a wide variety of cell types but exhibit an increased tumorigenic potential and lack target specificity when used for cell transplantation in the injured nervous system. The use of neural progenitor cells (NPCs) that have already undergone linage restriction reduces concerns about aberrant (non-neuroglial) phenotypes in the fully differentiated daughter cells, although their ability to generate a variety of surviving neuronal phenotypes is less certain. A successful transplantation of NPCs in acute neurodegeneration requires their survival in an unfavourable environment characterized by complex conditions of ischemia-like syndrome and neuroinflammation. Adult NSCs and ES cells accumulate at spinal cord injury (SCI) site, improve functional recovery, and die within 3 weeks. In view of such results we aimed at isolating ischemia-resistant neural precursors (PM-NPCs), their transplantation favours axonal regeneration, formation of a rich neuropile at injury site, and permanent recovery of function in models of spinal cord injury and Parkinson’s disease. We have recently been paying attention also to autologous adult stem cell sources. The differentiation potential and the ease of their isolation have made the multipotent mesenchymal stem cells very important for the development of a vast range of clinical applications in regenerative medicine and many laboratory and clinical settings have focused their attention on their use and development. Here we report our findings on human adipose tissue-derived stem cells (hADSCs) obtained from micro-fractured Lipogems-derived adipose tissue. The use of such a device allows the successful establishment of hADSCs colonies even without liberase H1 treatment, and after cryopreservation at -80°C. Differently it is difficult to obtain hADSCs from cryopreserved lipoaspirate. The cell cycle analysis showed that 75% of cells are in G0/G1phase and 21% in S+G2/M, and only a marginal 0.2% apoptosis. No Chromosomal abnormalities. These hADSCs from either fresh or frozen lipogems preparations are bearer of typical mesenchymal markers at values above 90%, and express embryonic markers such as NANOG and OCT4 and neural markers such as nestin, neurod1, pax 6 and musashi. The superficial epitopes are maintained even when hADSCs were grown in culture after storage at -80°C. Their driven osteogenic and adipogenic differentiation in vitro yields hADSCs with finer intracellular micro-organelles and fat deposits are more numerous and smaller in size. We shall also present how these lipogems-derived hADSCs can transdifferentiate in vivo in specific lineages determined by site of transplantation and condition of application.
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