The Experimental Research Of The Separation, Culture And Differentiation Of The Neural Stem Cell In Vitro

The aim of this study was to investigate the effects of the Hepatic Growth Factor(HGF), Insulin Like Growth Factor-1 (IGF-1) and Brain Derivation Neurotrophic Factor(BDNF) on the differentiation of the neural stem cell(NSC), and to detect the dynamic expression change of the bHLH gene-MASH1, NeuroD and Neurogenin2(Ngn2) in the process of neuronal differentiation of the neural stem cell, and to explore some inner molecular mechanisms of the differentiation. Cells isolated from the 14-17d embryonic rat brain were grown in the serum-free culture fluid. As the neural stem cells were gained, they were separated into 2 groups: HGF, IGF-1, BDNF were added into the experimental group selectively, with the Fetal Bull Serum(FBS) only into the control group. All kinds of the cells we gained pre and after the differentiation of the neural stem cell were identified by immunocytochemistical staining. Direct calculation and flowcytometre were applied to count the percentage of the neurons. The expression of the endogenous bHLH gene-MASH1, NeuroD and Ngn2 at the 1, 3 and 5 days following the differentiation were observed by RT-PCR. It was manifested by the primary culture that there were the morphological change in the cells growed in the culture medium continuously, in the day 4, the clonesphere...