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目的比较小鼠胚胎脑皮质与中脑来源的神经干细胞在培养与分化方面的差异,为更好的研究和利用神经干细胞提供实验依据。方法无菌条件下分别分离鼠胚大脑皮质与腹侧中脑,经胰酶消化及机械吹打成单细胞后接种于含有B27、bFGF的DMEM/F12培养基,培养扩增;倒置显微镜观察比较生长状况;机械方法传代;10%血清接种分化;免疫荧光细胞化学方法染色鉴定神经干细胞及其子代细胞的分化方向,并作比较。结果小鼠胚胎脑皮质与中脑均存在神经干细胞,其免疫细胞化学鉴定均呈Nestin阳性,并都能分化为神经元和胶质细胞;但皮质部位所含神经干细胞明显多于中脑,也更宜成球;有血清条件下分化,皮质神经干细胞未见分化为酪氨酸羟化酶(TH)阳性神经元;中脑神经干细胞则可少量分化为TH阳性神经元结论同样条件下皮质神经干细胞比中脑神经干细胞更易增殖与培养,两者在有血清条件下分化为TH阳性神经元能力方面有差异。
Objective To compare the differences in culture and differentiation of mouse embryonic cerebral cortex and midbrain neural stem cells in order to provide experimental evidence for better research and utilization of neural stem cells. Methods The aortic cortex and ventral mesencephalic were isolated under aseptic conditions. The cells were inoculated into DMEM / F12 medium containing B27 and bFGF by trypsin digestion and mechanical beating into single cells. The cells were cultured and expanded. By inverted microscope, Growth condition, passage by mechanical method, differentiation in 10% serum, differentiation direction of neural stem cells and progeny cells by immunofluorescence staining and comparison. RESULTS: Neural stem cells existed in the cerebral cortex and midbrain of mouse embryos. Immunocytochemistry showed Nestin-positive and all differentiated into neurons and glial cells. However, the number of neural stem cells in the cortex was significantly higher than that in the midbrain, More into the ball; under the conditions of differentiation, no differentiation of cortical neural stem cells into tyrosine hydroxylase (TH) positive neurons; a small number of mesencephalic neural stem cells can differentiate into TH positive neurons Conclusions Under the same conditions, cortical nerve Stem cells are more likely to proliferate and culture than midbrain neural stem cells, both of which have differences in their ability to differentiate into TH-positive neurons under serum conditions.