Culture and Identification of Rat Embryo Neural Stem Cells In Vitro

Objective To explore the conditions of culturing rat embryo neural stem cells in vitro and their identification.Methods The neural stem cells were derived from the cortex tissue of rat embryo at the age of 14～16 days and proliferated steadily under the effect of epidermat growth factor(EGF),basic fibroblast growth factor(bFGF) and B27.The rat neural stem cell sphere differentiation was induced by 10% fetal calf serum.The rat neural stem cells and the differentiated neural stem cells were identified respectively by immunofluorescence including Nestin,neuron specific enolase(NSE),gilial fibrillary acidic protein(GFAP) and Galc-C.Results The cultured neural stem cells proliferated,and become the sphere.The rat neural stem cells after the culture and passage were proved by positive Nestin.The differentiated cells in the sphere could express specific antigens of neurons(positive NSE),astrocytes(positive GFAP)and oligodendrocyte(positive Galc-C).Conclusion With the help of growth factors and serum-free technique,the rat neural stem cells which have potency of multiple differentiations can successfully be cultured in vitro.