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为了探讨胚胎大鼠脊髓神经管神经干细胞体外培养、鉴定的方法,本实验在解剖显微镜下机械性分离11.5 d的胚胎大鼠脊髓神经管,并制备细胞悬液,无血清培养基培养。应用免疫荧光染色方法对原代和传代细胞进行巢蛋白(nestin)鉴定;加入胎牛血清诱导其分化后分别行神经元特异烯醇化酶(NSE)、胶质原纤维酸性蛋白(GFAP)和髓磷脂碱性蛋白(MBP)鉴定。结果显示:11.5 d胚胎大鼠脊髓神经管来源的神经干细胞经连续传代培养可形成较多克隆球,呈nestin免疫阳性;加入胎牛血清可诱导其分别向神经元、星形胶质细胞和少突胶质细胞分化。本实验结果提示,利用E11.5的胚胎大鼠脊髓神经管可成功培养出大量稳定增殖并有多向分化潜能的神经干细胞,可用于进一步的研究。
In order to investigate the method of culturing neural stem cells from embryonic rat spinal cord in vitro and in vitro, the neural tube of the spinal cord of embryonic rat was mechanically separated for 11.5 days under a dissecting microscope, and the cell suspension was prepared and cultured in serum-free medium. Immunofluorescence staining was used to identify nestin in primary and passage cells. Fetal bovine serum (NSE), glial fibrillary acidic protein (GFAP) and myelin Phospholipid basic protein (MBP) identification. The results showed that neural stem cells derived from the spinal cord of 11.5-day-old embryos were subcultured and subcultured to form more spheres, which were positive for nestin. Addition of fetal bovine serum to neurons, astrocytes and less Dendritic cell differentiation. The experimental results suggest that a large number of neural stem cells that can stably proliferate and have multidirectional differentiation potential can be successfully cultured using the spinal cord neural tube of E11.5 embryos and can be used for further research.