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The aim of this study was to investigate the culture conditions of skin-derived mesenchymal stem cell (sMSCs) and to explore a new cell source for central nervous system cell transplantation. The cells from skins of mice were primarily isolated and cultured in serum-free medium, and they were transferred into serum-containing medium after passaged 2, and the passaged cells were identified by immunocytochemistry and induced to differentiate into multiple lineages. The results indicated that a population of sMSCs could be isolated from skins, they could be maintained in vitro for extended periods with stable population doubling, and they were expanded as undifferentiated cells in culture for more than 10 passages, indicating their proliferative capacity. About 60% of sMSCs expressed vimentin and the majorities of these cells expressed fibronectin. They could differentiate into adipocytes, osteogenic cells and fibroblast-like cells, they could differentiate into neurons with a simple protocol, and almost 50-60% of these cells expressed neuron specific enolase (NSE) and neurofilament (NF); and the differentiated neurons showed typical complicated morphology of neurons. In conclusion, skin contains stem cells that are capable of multiple differentiation; they could be cultured in vitro for long time and could maintain their characteristics of stem cells, and they may represent an alternative autologous stem cell source for CNS cell transplantation.

Objective: To investigate the possibility of inducing adipose tissue-derived stromal cells to differentiate into neuron-like cells, and to explore a new cell source for central nervous system transplantation., Methods: beta-mercaptoethanol was adopted to induce the cells to differentiate; undifferentiated cells and differentiated cells were identified with immunocytochemistry., Results: A population of adipose tissue-derived stromal cells were isolated from adult rat adipose tissue; they were processed to obtain a fibroblast-like population of cells and could be maintained in vitro for extended periods with stable population doubling, and they were expanded as undifferentiated cells in culture for more than 10 passages, indicating their proliferative capacity. beta-mercaptoethanol induced the stem cells to express nestin, characteristic of neuronal precursor stem cells at early stage of differentiation, and at late stage they exhibited a neuronal phenotype, expressing neuron-specific enolase (NSE) and neurofilament(NF); with an optimal differentiation protocol, almost 60%-85% of the cells expressed NSE and NF., Conclusion: The data support the hypothesis that adult adipose tissue contains stem cells capable of differentiating into neurons.

Objectives: To assess the culture and differentiation of neural stem cells in embryonic mice and set up a basis for further research in to neural stem cells., Methods: Embryonic cortices of mice were dissociated and single cell suspensions were achieved by mechanical methods in sterile conditions, and cells were seeded in uncoated plate in N2 medium. The cells were passaged by mechanical methods, frozen and thawed by general procedure. They were identified by immunocytochemical techniques., Results: Neural stem cells from embryonic mice were successfully cultured forming typical neurospheres in suspension. Neurons, astrocytes and oligodendrocytes were differentiated from neural stem cells, with a ratio of 7%, 85% - 90% and 2% - 4% respectively., Conclusions: Neural stem cells, which can be cultured and passaged steadily in vitro and they are the ideal cell sources for cell transplantation and gene therapy.