神经上皮干细胞的分离培养及其体外分化特性的观察  

ISOLATION AND CULTURE OF NEUROEPITHELIAL STEM CELLS AND OBSERVATION OF THEIR DIFFERENTIATION CHARACTERISTICS

作　　者：赵舒武[1] 高英茂[1] 张晓丽[1] 邴鲁军[1] 

机构地区：[1]山东大学医学院组织胚胎学教研室,济南250012

出　　处：《中国组织化学与细胞化学杂志》2005年第6期671-674,共4页Chinese Journal of Histochemistry and Cytochemistry

基　　金：山东省自然科学基金(01BB1DADA1)

摘　　要：目的探讨大鼠胚胎神经管神经上皮干细胞的分离培养条件,并观察其在体外的分化特性。方法采用显微解剖、机械吹打、无血清悬浮培养方法分离培养神经上皮干细胞,采用巢蛋白(nestin)免疫细胞化学染色技术检测神经上皮干细胞,用NSE和GFAP免疫组化染色检测并计数神经细胞和神经胶质细胞。结果大鼠胚胎神经管神经上皮干细胞在无血清培养基中可形成大量呈nestin抗原阳性细胞构成的神经球,经传代有血清培养后分化为NSE阳性和GFAP阳性细胞,其中NSE阳性细胞占细胞总数的47.7%,GFAP阳性细胞占细胞总数的39.8%。结论胎鼠神经管神经上皮干细胞在无血清培养中可增殖和传代,在有血清培养中可分化为神经细胞和神经胶质细胞,两者之比为47.7∶39.8。Objective To explore the isolation and culture conditions of neuroepithelial stem cells from neural tube of embryonic rats and to observe their differentiation characteristics in vitro. Methods The neuroepithelial stem cells were isolated and cultured by microdissection, mechanical blowing and serum-free suspension culture. The expression of nestin antigen, NSE and GFAP were identified by immunohistochemistry, and the number of NSE-positive cells and GFAP-positive cells were dividedly counted. Results In the serum-free suspension medium, neurospheres that consisted of a great number of nestin-postive cells could be obtained, and in the serum medium they differentiated into NSE-postive and GFAP-positive cells. The number of NSE-positive cells was 47.7% and that of GFAP-positive cells was 39.8%. Conclusion Neuroepithelial stem cells from neural tube of embryonic rats can be isolated, cultured and passaged in the serum-free medium, and the ratio of nerve cells to neuroglial cells in the serum medium is 47.7 ： 39. 8.

关 键 词：神经上皮干细胞 分离 培养 分化 大鼠 

分 类 号：R322.8[医药卫生—人体解剖和组织胚胎学]