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目的 以体外诱导生成的人神经干细胞 (NSCs)为模型 ,研究甲状腺激素 (TH)对其分化的影响。 方法 无菌状态下取因治疗目的而终止妊娠的胎龄 10～ 2 2周的人胎大脑半球 ,机械法分散细胞后 ,以 10 5个 ml细胞密度接种于含N 2配方的DMEM/F12培养基中 ,同时添加表皮生长因子 (EGF ,2 0 μg L)和 /或碱性成纤维生长因子(bFGF ,2 0 μg L)。采用自然分化以及T3诱导的方法诱导分化 ,免疫组织化学方法鉴定分化后的细胞类型 ,抗体分别为NF 2 0 0、GFAP、Gal C ,并采用抗MBP、O4、A2B5抗体鉴别少突胶质细胞发育的不同阶段。 结果 T3有助于向胶质细胞方向发展 ,包括少突与星形胶质细胞 ,MPB阳性细胞比例超过 80 % ,尤其在EGF +T3组 ,这种现象更为突出。 结论 NSCs的定时分化是内外源信号共同作用的结果。TH就是这样一种信号 ,激活“生物钟”机制的效应组件 ,在有限次的分化后诱导NSCs分化为少突胶质细胞
Objective To investigate the effects of thyroid hormone (TH) on the differentiation of human neural stem cells (NSCs) induced by in vitro culture. Methods Human embryonic brain hemispheres of 10 ~ 22 weeks gestational age and gestational age were aseptically removed. The cells were mechanically dispersed in DMEM / F12 medium containing N 2 at a density of 105 cells While epidermal growth factor (EGF, 20 μg L) and / or basic fibroblast growth factor (bFGF, 20 μg L) were added simultaneously. Differentiation was induced by natural differentiation and T3 induction, and the differentiated cell types were identified by immunohistochemistry. The antibodies were NF 2 0 0, GFAP and Gal C, respectively, and anti-MBP, O4 and A2B5 antibodies were used to identify oligodendrocytes Different stages of development. Results T3 promoted glial cells, including oligodendrocytes and astrocytes, with more than 80% of MPB positive cells, especially in EGF + T3 group. Conclusion The timing differentiation of NSCs is the result of the interaction of internal and external signals. TH is a signal that activates the effector component of the “biological clock” mechanism to induce the differentiation of NSCs into oligodendrocytes after a limited number of differentiations