[Comparisons among different methods of culturing neural stem cells isolated from human fetal cortex]

Neural stem cells (NSCs) are proved to be promising cell sources for gene therapy and cell therapy. To pursue optimal conditions for the isolation and culture of neural stem cells residing in human fetal cortex,the cortical tissue was dissociated mechanically and digested with various enzymes. Short-term trypsin digestion combined with pipette dissociation proved to be the suitable method of isolating human NSCs derived from the fetal cortex. Furthermore,DMEM/F12 medium was superior to the neurobasal medium in the aspect of clonal formation. In repeated dissociation experiments,it was found that accutase, instead of trypsin,endowed the NSCs with better growth and efficient neurosphere formation.