人胚中脑神经干细胞的分离、增殖及分化的研究

Isolation, proliferation and differentiation of neural stem cells from ventral mesencephalon of human fetal

目的 探讨人胚中脑腹侧神经干细胞的分离、增殖条件及分化方向。方法 应用表皮生长因子（EGF）和成纤维细胞生长因子-2（FGF-2）使人胚中脑腹侧幼稚细胞维持在未分化状态并促进其增殖;通过克隆分析、细胞增殖曲线及流式细胞仪检测培养细胞的增殖能力;应用免疫组织化学方法鉴定这些细胞的分化方向。结果 该细胞群具有一定的增殖能力,在撤除生长因子后至少能分化为神经元和星形胶质细胞,但未检测到多巴胺能神经元。结论 从人胚中脑腹侧分离出的幼稚细胞具有一定的增殖和自我更新能力,有多向分化潜能,具备中枢神经系统干细胞的一般特征。

Objective To study the isolation, proliferation and differentiation of the neural stem cells from the ventral mesencephalon of human fetal. Methods The immature cells derived from the ventral mesencepalon of human fetal were prevented from differentiating and prompted to proliferate with epidermal growth factor (EGF) and fibroblast growth factor (FGF-2). The proliferative capacity of these cells was estimated by cloning analysis, cell proliferative curves and flow cytometry and their differentiation were evaluated by inimunohistochemistry stainning. Results These neural stem cells had certain capacity of proliferating and renewing. They generated at least two kinds of mature cells of the central nervous system, including neurons and glia. Tyrosine hydroxylase (TH)-postive cells among these neurons were not detected. Conclusion Cells isolated from the ventral mesecephalon of human fetal have the common features of neural stem cells.