去上皮羊膜上培养兔角膜缘干细胞的实验研究

目的探讨去上皮羊膜上培养兔角膜缘干细胞的可行性。方法取小块兔角膜缘组织,采用组织块 细胞悬液共培养,待细胞长至对数生长期,分别传代于去上皮羊膜和完整羊膜,甲基偶氮蓝比色(MTT)法对比两组细胞增殖率,并行AE_5免疫鉴定。结果原代细胞接种7d左右,生长至对数生长期,细胞融合成单层。去上皮羊膜组细胞传代培养效果明显优于完整羊膜组。AE_5单克隆抗体染色强阳性。结论去上皮羊膜上培养出的兔角膜缘组织既有角膜上皮特性,又具有强的增殖潜力。

Culturing Rabbits' Limbal Stem Cells on Denuded Amniotic Membrane

Objective To evaluate the effectness of method which limbal stem cells were cultivated on denuded amniotic membrane.Methods Limbal comeae tissues of rabbit were cultured with limbal stem scrap tissues and cells suspension in vitro,and then drawn the cell growth curve of the limbal stem ceils.The cells were passaged on the intact and denuded amniotic membrance.MMT was used to detect the growth rate of two groups.The cul- tured cells were identified by AE_5 immunohistochemical staining.Results The cultivated rabbit limbal stem cells grew well in vitro,and achieved exponential phase of growth in seven days.The difference of MMT between the two groups was statistically significant(P<0.05).The culturing effectness of the denuded group was better than that of the intact group.The AE_5 immunohistochemical stain revealed the structural characteristic of normal comeal epithelium.Conclusion The rabbit limbal stem cells on denuded amniotic membrane not only have the character comeal epithelia but aslo have the high ability of proliferation.