神经干细胞与乳鼠基底膜共培养后分化为毛细胞样细胞的研究  

Differentiation of neural stem cells into hair cell-like cells by co-culturing with basilar membrane of newborn rats

作　　者：陈请国[1] 付勇[1] 王显红[2] 吴小慧[3] 谢静[1] 李哲文[1] 龚树生[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院耳鼻咽喉科,武汉430022 [2]咸宁学院附属第一医院耳鼻咽喉科,咸宁437100 [3]华中科技大学同济医学院附属协和医院检验科,武汉430022

出　　处：《中华耳科学杂志》2008年第2期216-220,共5页Chinese Journal of Otology

基　　金：国家自然科学基金资助项目(30672308)

摘　　要：目的探讨体外培养和鉴定胚胎大鼠神经干细胞及诱导其分化为毛细胞样细胞。方法从SD系胚鼠大脑分离神经干细胞,在无血清培养基中培养。取乳鼠基底膜与神经干细胞一起培养,14～21天后通过免疫荧光和免疫组化法检测毛细胞标志物myosin VIIa和calretinin。结果培养的神经干细胞胞体透亮,折光性好,Nestin鉴定阳性。诱导分化后的细胞免疫荧光和免疫组化示myosin VIIa和calretinin阳性。结论无血清条件下能培养出活性很好的神经干细胞,乳鼠基底膜能引导其朝毛细胞方向分化。Objective To investigate the method for culturing and identifying neural stem cells （NSCs） from rat embryos in vitro and inducing them to differentiate into hair cell-like cells. Methods NSCs were isolated from the brain of Sprague-Dawley rat embryos and cultured in serum-free conditional medium. The basilar membrane was isolated from post natal day 0 - day 3 rats and cultured in DMEM/F12 medium containing 10% fetal bovine serum, Then NSCs were co-cultured with basilar membrane for 14-21 days. The expression of hair cell markers （myosin VIIa and calretinin） was detected by immunocytochemical analyses. Results The cell bodies of NSCs were transparent with good refraction and marked by stem cell marker nestin. After differentiation, the cells were immunoreactive for myosin Ⅶa and calretinin, the hair cell marker. Conclusion NSCs grew well in serum-free conditional medium and they could be induced to differentiate into hair cell-like cells when co-cultured with basilar membrane from newborn rats.

关 键 词：神经干细胞 基底膜 分化 毛细胞样细胞. 

分 类 号：R322.923[医药卫生—人体解剖和组织胚胎学] R329.29[医药卫生—基础医学]