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目的了解角膜缘干细胞体外培养的增殖分化规律。方法组织块法培养细胞，测定细胞克隆形成率（ＣＦＥ），免疫荧光染色检测干细胞表达角质蛋白Ｋ３的状况。结果原代培养２１天左右细胞生长达到饱和，传第１代７～１０天形成单层，ＣＦＥ为９．５２％±４．９７％；传第２代７天，ＣＦＥ为４．２５％±２．１０％（Ｐ＜０．０１）。正常角膜缘基底细胞不表达角质蛋白Ｋ３；原代培养的干细胞亦不表达Ｋ３，传第１代细胞有部分表达。结论人角膜干细胞位于角膜缘基底部，培养的角膜缘干细胞早期具有较高的增殖力并保持干细胞的分化特性。
Objective To investigate the proliferation and differentiation of limbal stem cells cultured in vitro. Methods The cells were cultured by tissue block method and the rate of cell clone formation (CFE) was measured. The expression of keratin K3 in stem cells was detected by immunofluorescence staining. RESULTS: Primary cultured cells grew to saturation at 21 days. The first passage formed a monolayer from 7 to 10 days with a CFE of 9.52% ± 4.97%. On the second passage, CFE was 4.25% ± 2.10% (P <0.01). Normal corneal basal cells do not express keratin K3; primary cultured stem cells do not express K3, pass the first generation of cells are partially expressed. Conclusion Human corneal stem cells are located at the base of the limbus. The cultured limbal stem cells have higher proliferative capacity and maintain the differentiation characteristics of stem cells.