全反式维甲酸对小鼠胚胎不同阶段神经干细胞诱导作用研究

目的:探讨全反式维甲酸对小鼠胚胎不同阶段神经干细胞(NSCs)的诱导分化情况.方法:分离孕12.5d及15.5d的胚胎小鼠脑皮质.取第3代NSCs,用含1μmol·L-1的全反式维甲酸在体外诱导小鼠胚胎不同阶段的NSCs.诱导5d后,通过神经元微管相关蛋白2(MAP2)免疫荧光染色和Westernblots检测NSCs分化为神经元的比例.结果:与对照组相比,全反式维甲酸可明显提高神经元分化的比例.E12.5干细胞和E15.5胚胎干细胞分化为神经元的比例分别为30%±1.47%和16.21%±1.36%.结论全反式维甲酸具有显著的促神经干细胞分化成神经元的效应,并对胚胎不同阶段神经干细胞的诱导作用有所不同.

Experimental Study on the Differentiation of Mice Neural Stem Cells in Different Stages of Embryo into Neuron with All-Trans-Retinoic Acid (ATRA)

Objective:To investigate the differentiation of mice neural stem cells (NSCs) in different stages of embryo into neuron with all-trans-retinoic acid (ATRA). Methods:NSCs were separated from cerebral cortex of mouse embryos at E13.5 and E15.5. After 5 days in vitro,neurospheres were passaged. The cells of the 3rd passage were induced with ATRA. After 5 days,the capacity of NSCs differentiation toward neurons was determined by immunofluorescence double-labelling and western blot. Results: The inducing differentiation rates of NSCs have improved greatly by ATRA compared with control (P <0. 01). The differentiation ratio of neurons induced by ATRA in NSCs of E13.5 and NSCs of E15.5 was 30%±1.47%,and 16.21%±1.36% respectively after cultured 5 days,whereas the differentiation ratio of neurons was 12%±1.67% and 6.21%±1.12% respectively in control group. Conclusion: ATRA treatment can be induced to differentiation of NSCs into neurons. The NSCs from mouse embryos at E13.5 have more significantly promoted and can be induced to differentiate into neurons with ATRA in vitro compared with to the NSCs from mouse embryos at E15.5.