摘要
目的 :探讨体外诱导小鼠胚胎干细胞 (ESCs)分化为神经干细胞 (NSCs)的可能性。方法 :ESCs经胚胎体阶段 ,以视黄酸及视网膜m櫣ｌｌｅｒ细胞诱导 ,在NSCs选择性培养基中筛选培养扩增 7d ,观察形态变化 ,采用RT-PCR法检测nestin、glutaminase、Brn - 3基因表达及免疫细胞化学法检测nestin等NSCs特异性标志物 ,并对其扩增及分化能力进行观察。结果 :ESCs经初级诱导 ,NSCs选择性培养基筛选培养 7d后 ,形成大量神经球样结构 ,可扩增传代。绝大部分神经球样结构呈nestin抗原阳性 ,整合BrdU ,表达nestin、glutaminase、Brn - 3基因 ,且可分化为神经元及神经胶质样细胞。结论 :视黄酸及m櫣ｌｌｅｒ细胞诱导的ＥＳＣｓ,经选择性培养基筛选培养可获得NSCs 。
AIM:To explore the feasibility of inducing mouse embryonic stem cells into neural stem cells in vitro. METHODS: Embryonic body induced by retinoic acid and retinal müller cells were selected in neural stem cell-defined medium for 7 days, and the morphological changes were observed. The selected cells were stained immunocytochemically with anti-nestin, anti-BrdU antibodies, and their ability of expansion and differentiation were analyzed. RESULTS:Large amounts of neurospheres were derived from embryonic body in the selected medium on the 7th day, which could be passaged and differentiated, stained positive with nestin and BrdU, and expressed nestin, glutaminase and Brn-3 genes. CONCLUSION:Neural stem cells could be derived from embryonic body induced by RA and müller cells in the selected medium, which would offer an alternative to treat neuropathy such as glaucoma and retinal degeneration in the future.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2003年第3期289-292,T001,共5页
Chinese Journal of Pathophysiology
基金
国家重点基础研究发展规划项目 (973 )(G19990 5 43 0 1)
国家自然科学基金资助项目(No.3 0 1710 0 1
No .3 0 0 0 0 187)
广东省自然科学基金资助项目 (No .990 0 92
No.0 10 746)
