力学加载对胚胎神经干细胞骨架的调节作用

Effects of mechanical stimulation on cytoskeleton of neural stem cells from embryo of mouse in vitro

目的观察机械力学加载对体外培养的胚胎神经干细胞细胞骨架的调节作用。方法取妊娠13．5d昆明小鼠分离胚胎大脑中的神经干细胞，用添加碱性成纤维细胞生长因子的无血清培养基进行体外培养，在培养过程中对细胞进行力学加载，免疫细胞化学检测细胞表面抗原，用激光共聚焦显微镜观察不同组别的细胞骨架并进行定量分析。结果力学加载培养细胞和非加载细胞均呈100％巢蛋白阳性，力学加载组的细胞骨架较对照组清晰，荧光强度定量[（38.08±2．22）％]高于对照组[（25．19±2．44）％]，力学加载组与对照组的荧光强度差异有统计学意义（P〈0．05）。结论力学加载可以影响胚胎神经干细胞的骨架重排，从而可能影响细胞的功能和形态。

Objective To explore the effect of mechanical stimulation on the eytoskeleton of cul- tured neural stem cells from embryo. Methods Neural stem cells were isolated from cerebra of embryos of mouse at the gestational day 13.5. The harvested cells were euhured in serum-free media containing basic fibroblast growth factor （bFGF）, cyclic mechanical stretch was applied on neural stem cells by Flexereell 4000TM, the antigen on the cell surface was detected by using immunocytochemistry, and the cytoskeleton of the ceils was observed under the laser scanning confocal microscopy. Results About 100% of the differ- entiated ceils subject to cyclic mechanical stretch and controls were positive for nidogen. The cytoskeleton fluorescence intensity of the cells subject to stretch was （38.08 ± 2. 22） %, and that in the control group was （25.19 ± 2. 44） % （P 〈 0.05 ）. Conclusion Cyclic mechanical stretch can affect the cytoskeleton of neural stem ceils from embryo in vitro, which may regulate the physiology of the neural stem cells.