Extablishment of adult neural stem cell culture

Objective To investigate the method of adult neural stem cell (NSC) culture in vitro.Methods The neural stem cells were derived from the brain tissue of 6-week-age rats and proliferated steadily under the effect of neural stem cell culture fluid (DMEM/F12 with 1% N2, 2% B27, 20 μg/LEGF and 20 μg/L bFGF). Neural stem cell ifferentiation was induced by 10% fetal calf serum. The change of cell morphology was observed under an inverted microscope. The rat neural stem cells and the differentiated neural stem cells were identified respectively by flow cytometry including Nestin, neuron specific enolase (NSE) and Galc-C. Results The cells derived from the brain tissue could be subcultured in vitro. The rate of positive Nestin cells was 97.6%. After induction by 10% fetal calf serum, these cells were positive for NSE and Galc-C. Conclusion By using fetal calf serum-free neural stem cell culture fluid, the rat adult neural stem cells having potential of multiple differentiations can successfully be cultured in vitro. Key words: Adult neural stem cells; Culture in vitro