Cultivation,proliferation and differentiation of human embryonic neural stem cells in vitro

Objective To establish the methods of isolation,cultivation and in vitro differentiation of neural stem cells and to observe the characteristics of proliferation and differentiation of neural stem cells.Methods The serum free culturing technology was used to isolate,culture and pass neural stem cells from embryonic human hippocampus.Immunocytochemistry,flow cytometry and cell growth curve were used to identify the neural stem cells and their potentiality of differentiation and proliferation.Results The neurospheres isolated from embryonic human hippocampus possess the ability to proliferate,self renew and passage.Nestin positive cells can be found in the neurospheres.After attachment they can differentiate into GFAP and NSE positive cells.Conclusion The methods to isolate and culture neural stem cells from embryonic human hippocampus are established.The cultured neurospheres possess the capability of proliferation,self renew and multipotentiality.The neurospheres that express nestin are the neural stem cells.It can be used to study on cell transplantion.