论文部分内容阅读
目的探讨分离培养人脑神经干细胞的方法及条件。方法用胰蛋白酶消化法从人胚脑中分离单个细胞,用无血清培养基进行体外培养,bFGF和EGF刺激细胞生长,用血清诱导其分化;采用免疫荧光细胞化学染色方法检测培养的细胞神经巢蛋白(Nestin)抗原和分化后成熟神经细胞抗原神经元特异性烯醇化酶(NSE)的表达。结果从胚龄16周的新鲜人胚脑中成功分离出神经干细胞。该细胞可在体外培养条件下传代培养,并表达神经干细胞标志Nestin;分化后的细胞主要表达成熟神经细胞标志NSE。结论在体外培养条件下可从人胚脑中分离培养出神经干细胞。
Objective To investigate the methods and conditions for the isolation and culture of human neural stem cells. Methods Single cells were isolated from human embryonic brain by trypsin digestion, cultured in serum-free medium, bFGF and EGF stimulated the growth of cells, and induced differentiation by serum. Immunofluorescence staining was used to detect the cell nests Nestin antigen and mature neuronal cell antigen (NSE) after differentiation. Results Neural stem cells were successfully isolated from fresh human embryonic brain of 16 weeks old. The cells can be subcultured in vitro and expressed neural stem cell marker Nestin; the differentiated cells mainly express mature neuronal cell marker NSE. Conclusion Neural stem cells can be isolated and cultured from human embryonic brain under in vitro culture conditions.