| BackgroundGlioma cells and neural stem cells may have an intrinsic link between. The present study showed that glioma cells secrete various cytokines, can promote the proliferation and migration of NSCs. However, the mechanism of the differentiation and Migration of neural stem cells Induced by glioma cell is unclear. The subject our group study the phenomena of migration and differentiation of neural stem cells induced by glioma cells inn vivo and In vitro.In vivo:producting a model rat with glioma, and transplant the neural stem cells derived from GFP (green fluorescent protein) transgenic mice into the Parahippocampal area of SD rats with glioma, we observe that cell differentiation of the GFP-positive cells into neurons and astrocytes expressed specific antigens by flow cytometry and immunohistochemistry, and track the survival and the differentiation of neural stem cellsExperiment 1 Isolation,culture and identification of neural stem cells of GFP transgenic micePurposes:separation, culture and identification of NSCs of GFP transgenic mice and provide neural stem cells for the transplantation experment of NSCs and the experment in vitro.Methods:â‘ isolated GFP transgenic fetal mouse hippocampus, wind and percussion with a mechanical suction into single cell suspension. The method includes the B27, basic fibroblast growth factor (bFGF) and epidermal growth factor (EGF) in serum-free culture medium.â‘¡The Nestin antibody on primary and passaged cells specific identification.â‘¢with fetal bovine serum induced differentiation, the differentiated cells were observed under fluorescence microscope morphology and GFP expression. Using NeuN, GEAp and 04 antibody respectively differentiated neurons, astrocytes and glial cells of small sticks of specific identification.Results:â‘ from GFP transgenic fetal rat hippocampus cells isolated form of suspended growth, can be formed typical neurospheres, in culture, and continuous subculture in the formation of cloning, and primary and passage cells were positive for GFP expression.â‘¡immunocytochemistry showed primary and passaged cells were Nestin positive.â‘¢differentiated cells induced by serum can be separately expressed neurons, astrocytes and Oligodendroglioma specific antigen NeuN, GEAp and 04, and the differentiated cells were positive for GFP expression.Conclusion: GFP transgenic embryo isolated and cultured rat hippocampus success out NSCS. Nestin-positive cells cultured with proliferation and self-renewal and to the neurons and glial cell differentiation potential, and can be stably expressed GFP, NSCs transplanted into the brain can be used as experimental study of donor cells.Experiments two chemotaxis and differentiation experiment of neural stem cells in vivoPurposes:validation transplantation GFP transgenic neural stem cells in the parental animals ability to survive, to tumor tissue chemotaxis and to study the neural stem cells contained tumor rats transplanted tumor growing areas of regional and relative distribution of differentiation.Methods:First established glioma model, model after successful, stem cell transplantation in 14 days group,21 days group and sham group were killed at the relevant time experimental animals, drawn fresh sterile rat brain tumor growing areas and the positioning of neural stem cell transplantation area, collecting center mark about 4* 4* 4mm3 size of brain tissue, brain tissue homogenate, filtration, breaking Remover rupture of membranes, the attainment homogenate, rabbit anti-mouse nestin antibody (1:1), neun antibody (1:1), labeled cell homogenate, with Alsace red fluorescent secondary antibodies labeled (goat anti-rabbit 1:1), flow cytometry on a comparison of different neural regions stem cell differentiation.Results:By flow cytometry using fluorescence labeled antibody associated antibodies observed in GFP transgenic mice transplanted neural stem cells in the living state, the distribution and differentiation.Conclusion:The transplantation of neural stem cells in vivo with glioma cells to chemotaxis and proliferation; relative to the original graft area, migration of nerve cells surrounding tumor clear trend in the neuronal differentiation, but differentiated into neurons value-added trends in 7-14 days, gradually slowing down.
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