力学加载对胚胎神经干细胞分化的影响

Effects of mechanical stimulation on differentiation of neural stem cells from embryo of mice in vitro

目的观察机械力学加载对胚胎神经干细胞（NSC）体外培养和分化的影响。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（bFGF）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及细胞分化表面抗原。结果力学加载培养细胞和非加载细胞均呈100％巢蛋白阳性，分化良好，对照组非力学加载分化细胞中（10．9±4．7）％细胞呈Tubulin阳性，力学加载4h后的胚胎神经干细胞的阳性率为（20．5±3．4）％，力学加载后的培养细胞和非加载培养细胞的分化率差异有统计学意义（P〈0．05）。结论力学加载对体外培养的胚胎神经干细胞的分化有促进作用。

Objective To explore the effect of mechanical stimulation on the differentiation of neural stem cells （NSC） from embryos in vitro. Methods Neural stem cells were isolated from cerebra of mouse embryos with pregnant durations of 13.5 days. The harvested cells were cultured in serum-free media containing basic fibroblast growth factor （ bFGF）, and cyclic mechanical stretch was applied on neural stem cells by Flexercell 4000TM, The antigen on the cell surface was detected by using immunocytochemistry. The differentiation rate of the ceils was observed under an inverted microscopy. Neural stem cells were identified by using immunocytochemistry. Results About 100% of the differentiated cells with or without cyclic mechanical stretch showed positive nidogen. The percentage of Tubulin positive cells in stretch group and control group was （20. 5 ± 3.4） % and （ 10. 9 ± 4. 7 ） %. The differentiation rate of the ceils in stretch group was increased significantly as compared with control group （P 〈 0. 05 ）. Conclusion Intermittent loading can affect the differentiation of neural stem ceils from embryos in vitro. It can promote neural stem ceils to differentiate into neurons.