| Objective: To establish the method of culture neural stem cells(NSCs)isolated from olfactory bulb of neonatal rats in vitro and to observe the proliferation of NSCs. To investigate the proliferation and differentiation characters of NSCs by observing the cell morphous and analysising the immunofluorescence result of NSCs from the olfactory bulb of rat at different stage in vitro. By using fetal bovine serum of different density, IGF-I and BDNF to induced NSCs of olfactory bulb, to explore the factors that spur the NSCs to differentiate and to pave the way for the study in future.Methods: NSCs from olfactory bulb of neonatal rats were isolated, cultured and expanded by neurosphere formation using serum-free medium containing growth factors, bFGF and EGF. The cells were passaged continuously by dissociating mechanically in order to harvest them in bulk. The cell morphous was observed under phase-contrast microscope and electronmicroscope. Immunocytochemistry was performed to detect the expression of Nestin. The cell proliferation was confirmed by 3(4,5dimethythiazol-2-yl)-2,5-diphenyl tetrazolium bromid (MTT) colorimetric assay... |
