ABSTRACT
Objective To investigate the effects of passaging on the proliferation,differentiation and apoptosis of neural stem cells.Methods Neural stem cells both from prime culture(P0) and twice-passaged culture(P2) were tested by enzyme immunoassay for proliferation and fluorescent activated cell sorting for apoptosis by Annexin V-FITC and propidium iodide.After being enduced to differentiation for 7 days,they were detected by the neuronal marker MAP2 and the astrocyte marker GFAP with nuclei staining by Hoechst 33342 additionally.The ratios of MAP2+/Hoechst+ and GFAP+/Hoechst+ were calculated.Results The absorbance of NSCs from P0 and P2 is 1.1899?0.0485(n=13)and 0.3526?0.1053(n=11)(P=0.000).Analyses of apoptosis using cytoflurometry are 40.91%?12.74% for P0(n=8)and 24.31%?13.01% for P2(n=9,P=0.018).The ratios of MAP2+/Hoechst+ and GFAP+/Hoechst+ of NSCs from P0 were 58.81%?9.69%(n=10)和22.85%?4.36%(n=10) while the ones for P2 were 27.84%?5.70%(n=10)and 49.32%?6.95%(n=10,P=0.000).Conclusions By passaging the proliferation and apoptosis of NSCs decreases and they differentiate into less neurons and more astrocytes.