大鼠胚胎脑组织神经干细胞的培养和鉴定

Culture and Identification of Neural Stem Cells from Brain Tissue of Rat Embryo

目的探讨从不同胎龄的大鼠脑组织中分离,培养神经干细胞(NSC)并对其鉴定,了解生物特性。方法通过采用机械分离和消化分离相结合的方法分离不同胎龄大鼠脑NSC。在无血清DMEM/F12(含20ng/m lbFGF,20ng/m lEGF及B27辅助培养液)中培养、传代和鉴定。诱导分化后采用SABC法对分化的细胞进行神经元特异烯醇化酶(NSE)、胶质纤维酸性蛋白(GFAP)检测作细胞鉴定。结果从不同胎龄的胎鼠脑组织中成功培养出神经干细胞,胎龄为12.5天的胎鼠提取的神经干细胞集落最多,在上述条件下培养及传代的细胞不断分裂增殖,形成悬浮生长的呈巢素蛋白(nestin)阳性的神经球;用血清诱导分化为大量表达NSE阳性的神经元和GFAP阳性的星形胶质细胞。结论胎龄为12.5天胎鼠大脑皮质培养出的神经干细胞数量最多,可分化为神经元、神经胶质细胞及少突胶质细胞。

Objective To isolate,culture and idenfify neural stem cells （NSCs） from brain fissue of rat embryoes in different fetal age. Methods NSCs of different fetal age rats were isolated and cultured through the combination of mechanical and pancreatin digestant isolation. Neurospheres and differentiated cells were identified by immuncytochemisty. Results NSCs were incubated from the brain tissue of different fetal ages successfully with the most NSCs number for 12.5 days fetal rat comparing other fetal ages. Neurospberea formed by NSCs were nestin posifive, and furthermore differentiated into neuron specific enolase positive ceils and glial fibrillary acidic protein positive cells respectively. Conclusion The brain tissue of the different fetal age rat could be cultured into neural stem cells with the most number of ceils for 12.5 dags embryo,which furthermore differentiated into neurons and glial cells.