保存液中添加bFGF对中期保存角膜细胞活性的影响

A study of basic fibroblast growth factor on corneal cell viability in mid-term preserved medium in vitro

目的 :研究bPGF对中期保存角膜细胞活性的影响。方法 :配制保存液保存新西兰大白兔角膜 ,对照组为配制保存液保存 ,实验组为配制保存液加入bFGF(10ng/ml) ,两组在 30～ 34℃恒温条件下保存 16～ 18h后转入 4℃低温保存 ,在保存3、5、7d分别取保存角膜片行 30～ 34℃ 2h复温 ,台盼蓝 -茜素红联合染色检测角膜内皮细胞活性、电镜检测细胞超微结构和PCNA检测细胞增殖情况。结果 :加入bFGF实验组保存角膜 ,角膜水肿较轻、皱褶较少、透明度较高 ,角膜内皮细胞形态结构和活性较好 (P <0 .0 5 ) ;并且角膜上皮、基质和内皮层有PCNA阳性细胞 ,与对照组比较差异有显著性 (P <0 .0 5 )。结论 :bFGF在中期角膜保存中有促进细胞增殖、保持角膜细胞活性的作用 ,可应用于中期角膜保存。

Objective:To investigate the effect of basic fibroblast growth factor (bFGF) on corneal cell viability in mid-term preserved medium.Methods:prepared the medium which contained minimum essential medium (MEM),dextran (40000),HEPES and so on.New Zealand′s white rabbits′ corneas were preserved in the medium and the medium with bFGF(10ng·ml -1 ) at 30～34℃ for 16～18 hours,and then transmited to 4℃to be storaged.At 3rd,5th,7th day after preservation,the viability of corneal endothelial cells (CECs) were assessed by trypan blue-alizarin red stain;the ultrastructure of comea was observed by electron microscope.Corneal cell proliferation was monitored by PCNA immunohistochemistry.Results:The corneal edema and folds in the medium with bFGF are slight.The cornea transparency,cell shape and the cell viability in bFGF group are better than that in matched control group(P<0.05 ).PCNA-positive cells in corneal epithelial,stromal and endothelial layers in experimental groups were significantly more than that in matched control group(P<0.05) .Conclusion:The results indicated that bFGF play a significant role in stimulating cell proliferation and maintaining the structure and viability.It can be administered to corneal mid-term preserved medium.