新生大鼠海马神经干细胞的分离培养及分化研究

发布时间：2018-03-03 10:20

本文选题：海马　切入点：神经干细胞　出处：《生物医学工程研究》2016年04期 　论文类型：期刊论文

【摘要】：研究新生大鼠海马区脑组织中神经干细胞体外培养方法,为治疗神经系统疾病寻找合适的细胞来源。取新生SD大鼠的海马区脑组织,采用accutase结合机械分离法获取神经干细胞,在含有B-27、碱性成纤维生长因子和表皮生长因子的DMEM/F12无血清培养液中培养;Accutase酶消化后传代培养,取第3代细胞行抗巢蛋白免疫荧光染色鉴定并以含10%胎牛血清培养液诱导分化,神经元特异烯醇化酶和胶质纤维酸性蛋白免疫荧光染色检测NSCs向神经元及胶质细胞分化的能力。分离的新生大鼠海马区脑组织中细胞,在无血清培养液中形成大量的神经球,部分神经球出现融合及贴壁分化现象,细胞呈典型NSCs形态。经巢蛋白染色鉴定,大部分为阳性细胞。神经细胞球经含有胎牛血清培养液培养后,可分化为神经元特异烯醇化酶和胶质纤维酸性蛋白表达阳性的细胞。从新生大鼠海马组织分离培养的NSCs具有自我更新和增殖能力,在含胎牛血清培养液中具有向神经元和神经胶质细胞分化的潜能。
[Abstract]:In order to find a suitable cell source for the treatment of nervous system diseases, the neural stem cells in the hippocampus of newborn rats were cultured in vitro. Neural stem cells were obtained from the hippocampal brain tissues of newborn SD rats by accutase and mechanical separation. DMEM/F12 containing B-27, basic fibroblast growth factor and epidermal growth factor was cultured in serum-free medium after digestion and subculture. The third passage cells were identified by anti-nestin immunofluorescence staining and differentiated with 10% fetal bovine serum culture medium. Neuron-specific enolase and glial fibrillary acidic protein immunofluorescence staining were used to detect the ability of NSCs to differentiate into neurons and glial cells. Fusion and adherent differentiation were observed in some of the neurons, and the cells showed typical NSCs morphology. Most of the neurons were positive cells identified by nestin staining. The neuronal balls were cultured in the culture medium containing fetal bovine serum. NSCs isolated from hippocampal tissue of newborn rats has the ability to self-renew and proliferate, and can differentiate into neuron-specific enolase and glial fibrillary acidic protein (GFAP) positive cells. It has the potential to differentiate into neurons and glial cells in fetal bovine serum culture medium.
【作者单位】：江苏无锡市第三人民医院;
【分类号】：R651.2

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