神经生长因子(NGF)对兔角膜缘干细胞增殖的影响

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5×103个/m l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M TT法测定细胞增殖情况。结果:10ng/m l、50ng/m l、100ng/m l、200ng/m l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/m l组与对照组、50ng/m l组与200ng/m l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10~100 ng/m l,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。

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

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 cells 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.