RATIO OF NEURON AND GLIA DIFFERENTIATED FROM THE EMBRYONIC HUMAN NEURAL STEM CELLS AT DIFFERENT TIME IN VITRO / 解剖学报

ABSTRACT

Objective To compare the ratios of neuron and glia differentiated from the embryonic human neural stem cells(hNSCs) at different times in vitro. Methods The cells from hippocampus of aborted human embryo tissue(8-12 weeks after conception) were mechanically dissociated and cultured with DMEM/F 12 medium containing h-EGF,h-bFGF,h-LIF.The NSCs were induced to differentiate by 1%FBS and identified by Musashil,?-Ⅲ Tubulin,GFAP and Galc immuno-cytochemical staining at different times. Results The cell spheres were Musashil positive at the 12th hour after differentiation.On the 2nd day after differentiation,a few cells exhibited ?-Ⅲ Tubulin and GFAP immunoreactivities,but no Galc positive cells.The number of ?-Ⅲ Tubulin immuno-positive cells reached the highest on the 7(th) day and the percent was 48.2%.The number of GFAP immuno-positive cells increased from the 2nd to the 23rd day and the positive ratio reached 65.3%.A few Galc immuno-positive cells were detected with time passing.Conclusion 1% FBS may promote human NSCs to differentiate into three cell types of central nervous system,and the ratios of differentiated neuron and glia are significantly different at various times.