胚胎小鼠听皮层区域神经干细胞的分离培养及鉴定

摘要

目的 通过剖腹手术从C57BL/6胎鼠(El4左右)中获得听皮层区域组织,进行神经干细胞体外培养和分化鉴定,探索新的自体NSC移植治疗来源.方法 选取孕14天左右的C57BL/6小鼠,剖腹取出胎鼠,分离听皮层区域组织,在体外无血清培养得到NSC,用免疫荧光细胞染色法进行NSC特异性标记物(Nestin)、增殖能力(BrdU)及多向分化潜能的鉴定.结果 听皮层区域组织在体外通过无血清培养能够得到大量细胞球,经Nestin及Brdu鉴定为NSC球且具有较高增殖能力.NSC球在体外分化后产生β微管蛋白(β-TubulinⅢ)阳性的神经元以及胶质纤维酸性蛋白(GFAP)阳性的星形胶质细胞.结论 胎鼠听皮层区域在体外无血清培养可获得大量具有增殖能力和多向分化潜能NSCs,是自体NSCs移植治疗新的种子来源.%Objective To culture and identify the neural stem cells (NSCs) derived from auditory cortex in C57BI7 6 embryo mouse (E14 days) by laparotomy and search for a new source of NSCs for self-transplantation therapy. Methods Auditory cortex was obtained from embryo mouse acquired from C57BL/6 pregnancy mouse (E14days) by laparotomy. NSCs were isolated from auditory cortex and cultured in serum-free medium in vitro. Fluorescence immunocytochemis-try was carried out to examine the expression of NSCs marker (nestin), self-proliferation (Brdu), and the multipotential of differentiation. Results A large number of NSCs examined by Nestin could be harvested from Al in vitro in serum-free media. They expressed Brdu, showing their potential of self-proliferation. They were also able to differentiate into β-Tubu-lin Ⅲ positive neurons and GFAP positive astrocytes. Conclusion NSCs can be obtained from Al of embryo mouse by in vitro serum-free culture, with potentials of self-proliferation and multipotential of differentiation. This can be applied to self-transplantation therapy as a new source of NSCs.