The long-term culture and passaging of human neural stem cells

Objective To explore the culture conditions for human neural stem cells and to investigate the passaging method.Methods The cells from the embryonic human cortex were mechanically dissociated.N2 medium was adapted to culture the cells, bFGF and EGF were added to expand the cells, The cells were identified by immunocytochemistry.Results Neural stem cells from embryonic humans have been successfully cultured. They formed typical neurospheres in suspension,and the majorities of the cells expressed vimentin and Musashil,which were the markers for neural stem cells. The cultured cell could differentiated into neurons and astrocytes. The neural stem cells multiplied very slowly under the culture conditions, however the best expansion was achieved when the neurospheres were disected into several parts and the cell link was conserved when passaging.Conclusions Human neural stem cells could be cultured from embryonic brains.They could form the typical neurospheres in suspension in vitro, which may be potential source for transplantation in treating CNS disorders in humans.