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The Research of Isolation， Proliferation and Differentiation of Neural Stem Cells from the Newborn Rat Hippocampus in vitro
Bai Ruiying1, Zhang Zijuan2, Wang Yali1, Li Xinjuan1, Cheng Yuan1, Wei Linyu1, Xue Hong2*
1Department of Physiology and Neurobiology, Xinxiang Medical University, Xinxiang 453003, China; 2Basic Medical Sciences of Henan University of TCM, Zhengzhou 450008, China
Abstract: The experiment aimed to explore the feasibility of the isolation， proliferation and differentiation of neural stem cells (NSCs) from the hippocampus in vitro. NSCs were isolated sterilely from the hippocampus of newborn (24 h) SD rat， which were cultured in the serum-free medium and induced to differentiate by the fetal bovine serum induction medium. Immunofluorescence was performed to detect Nestin， NSE and GFAP in order to identify the cell types， respectively. And flow cytometry was used to determine proliferative activity of NSCs before and after induction. The results demonstrated that the cells isolated from the hippocampus had the ability of proliferation and could futher form clone spheres and express Nestin. In addition， the differentiated cells expressed NSE and GFAP， respectively. Meanwhile， the results from flow cytometry showed that NSCs were actively dividing and the percentage of cells in S+G2/M was (36.27±1.99)%. However， at development stages of differentiation (3， 7， 10 d)， the percentages of cells in S+G2/M were obviously decreased (P<0.05)， which were (26.39±1.10)%， (26.33±1.33)%， (24.54±1.12)%， respectively. In conclusion， neural stem cells exist in the hippocampus of newborn (24 h) rat， which have the ability of self-renewal and multipotent differentiation， and would be very useful for basic and clinic research.
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