大鼠海马神经干细胞的分离、培养及鉴定

南方医科大学学报 ›› 2004, Vol. 24 ›› Issue (10): 1143-1146.

大鼠海马神经干细胞的分离、培养及鉴定

郭洪波, 邹飞

南方医科大学热带卫生学系高温医学研究室, 广东, 广州510515

出版日期:2004-10-20 发布日期:2004-10-20
基金资助:
收稿日期:2004-2-10。
基金项目:国家自然科学基金(30371575)
作者简介:郭洪波(1971- ),女,南方医科大学在读博士研究生,电话:020-61640114-89124
通讯作者:邹飞,电话:020-61648301

Isolation, culture and identification of rat hippocampal neural stem cells

GUO Hong-bo, ZOU Fei

南方医科大学热带卫生学系高温医学研究室, 广东, 广州510515

Online:2004-10-20 Published:2004-10-20

摘要/Abstract

摘要： 目的探寻出生后1~3d和成年大鼠海马分离、培养并鉴定神经干细胞(NSC)的可行性。方法向培养基添加碱性成纤维细胞生长因子、脑源性神经营养因子及其他生长因子,采用单细胞克隆技术,分离出生后1~3d和成年SD大鼠海马组织并分别无血清培养;以免疫细胞化学方法对原代和传代培养形成的克隆细胞,以及其分化后的细胞进行鉴定。结果上述条件下培养可形成悬浮生长的表达巢蛋白的神经球,且具有连续克隆的能力;分化后可得到具有网状联系的、分别呈神经元特异性烯醇化酶阳性和胶质纤维酸性蛋白阳性的神经元及胶质细胞。结论新生及成年SD大鼠海马存在神经干细胞。

Abstract: Objective To explore the feasibility of the isolation,culture and identification of the neural stem cells originated from the hippocampi of neonatal rats (1-3 d) and adult rats respectively.Methods The neural stem cells were isolated from the hippocampi of neonatal and adult rats and cultured in serum-free medium containing basic fibroblast growth factor (bFGF),brain-derived neurotrophic factor (BDNF) and other growth factors using single-cell cloning technique. Immunochemistry was employed to identify the cloned cells and their daughter cells as well as mature neural cells differentiated from the cell clone. Results The floating neural spheres obtained by above culture procedures for isolated cells from the hippocampi of neonatal and adult rats were positive for nestin expression,with the potential for further cloning and capable of differentiation into neurons or glial cells,which formed reticular connection between the cells and expressed neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) respectively.Conclusion The hippocampi of neonatal and adult rats contain neural stem cells.

中图分类号:

R364
Q28

郭洪波, 邹飞. 大鼠海马神经干细胞的分离、培养及鉴定[J]. 南方医科大学学报, 2004, 24(10): 1143-1146.

GUO Hong-bo, ZOU Fei. Isolation, culture and identification of rat hippocampal neural stem cells[J]. Journal of Southern Medical University, 2004, 24(10): 1143-1146.

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