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目的建立人胚胎来源的神经干细胞体外培养方法,初步研究挫伤脑组织提取液(TBITE)对人胚神经干细胞(HNSCs)存活、增殖能力和分化的影响。方法10周龄左右的胎儿大脑皮层细胞体外培养,神经干细胞增殖2～3代后,剔除生长因子,分别加入不同浓度的脑外伤病人的挫伤脑组织提取液,培养2-3周后,免疫细胞化学方法鉴定细胞分化后的类型。用Laemmli凝胶电泳法分析培养液上清。结果成功获得以神经球形式生长的人胚神经干细胞,Nestin表达阳性;与空白对照相比,TBITE组中的神经干细胞有明显的增殖,并分化为神经细胞、星形胶质细胞和少突胶质细胞。随着TBITE含量的增加,星形胶质细胞的比例增加。结论脑外伤的局部微环境可促进神经干细胞增殖和分化。
Objective To establish a method for culturing human embryonic neural stem cells in vitro and to study the effects of TBITE on the survival, proliferation and differentiation of human embryonic neural stem cells (HNSCs). Methods Fetal cortex cells of 10 weeks old were cultured in vitro. After 2 ~ 3 generations of proliferation of neural stem cells, the growth factors were removed and the brain tissue extracts of contusion of different concentrations of traumatic brain injury were added respectively. After 2-3 weeks of culture, immune cells Chemical methods to identify the type of cell differentiation. The culture supernatant was analyzed by Laemmli gel electrophoresis. Results Neural stem cells were obtained in neurospheres, and Nestin expression was positive. Compared with the blank control group, neural stem cells in TBITE group proliferated and differentiated into neurons, astrocytes and oligodendrocytes Stromal cells. As the TBITE content increased, the proportion of astrocytes increased. Conclusion The local microenvironment of traumatic brain injury can promote the proliferation and differentiation of neural stem cells.