神经生长因子(NGF)对兔角膜缘干细胞增殖的影响

Effect of never growth factor on proliferation of rabbit corneal limbal stem cells

目的：探讨神经生长因子（NGF）对兔角膜缘干细胞增殖作用的影响及最佳有效浓度。方法：取成年大耳白兔角膜缘组织，应用组织块消化培养法进行原代培养，取第2代生长状态良好的干细胞，以5×10^3个／ml细胞浓度接种于96孔细胞培养板中，依次加入不同浓度的NGF．培养48h后，采用MTT法测定细胞增殖情况。结果：10ng／ml、50ng／ml、100ng／ml、200ng／ml不同浓度组的NGF均有促进角膜缘干细胞的增殖作用，与对照组比较差异均有显著性（P〈0．01）。1ng／ml组与对照组、50ng／ml组与200ng／ml组比较差异均无显著性（P〉0．05）。结论：NGF可有效促进培养角膜缘干细胞的生长，培养的适宜浓度为10～100ng／ml，为角膜缘干细胞移植患者术后及角膜外伤或溃疡时局部应用NGF提供了理论依据。

Objective： To investigate the effect of never growth factor （NGF） on proliferation of rabbit corneal limbal stem cells and its optimal dose in vitro. Methods：The rabbit limbal tissues were cultured by enzymatic digestion culture method. The second generation stem ceils were inoculated in 96 well plates at a density of 5000 cells per milliliter, treated with different concentrations of NGF. After 48 hours of culture, the cell proliferation was assessed by MTT assay. Results： The experimental groups at the concentrations of 10 ng/ml, 50 ng/ml, 100 ng/ml and 200 ng/ml had significantly higher cell viability than that of the control group （P 〈 0.01） , with no significant difference between the 1 ng/ml group and the control group, and also between the 50ng/ml group and the 200 ng/ml group （P 〉 0.05） . Conclusion： NGF enhances the proliferation of the stem cells in vitro effectively. The optimal concentration ranges from 10ng/ml to 100ng/ml. The experiment provides the theoretical basis for using NGF for surgical transplantation and for patients suffering from corneal trauma or ulcer.