不同部位角膜上皮细胞体外增殖性的对比研究

Comparative Study of Proliferation of Limbal, Peripheral and Central Corneal Epithelial Cells in Culture

为观察角膜缘、角膜周边及角膜中央部上皮细胞体外增殖的规律，确定角膜干细胞以选取体外培养角膜上皮细胞的部位。应用溴代脱氧尿嘧啶（Ｂｒｄｕ，胸腺嘧啶的类似物）掺入ＤＮＡ合成，抗－Ｂｒｄｕ单克隆免疫荧光抗体记标记细胞法，在流式细胞仪（ＦＡＣＳｔａｒｐｌｕｓ）上检测体外培养的每一代角膜中央部、周边部及角膜缘上皮细胞增殖的情况。结果显示从第一代开始，角膜缘角膜周边角膜中央部上皮细胞增殖率逐渐递减；角膜缘、角膜周边、角膜中央部细胞代数依次减少；原代角膜缘及角膜周边部上皮细胞增殖率经统计学处理，无显著性差异（Ｐ＞００５）；结论：（１）角膜缘部细胞增殖潜力大；（２）角膜缘上皮细胞群中存在一定数量的长周期细胞。因此，角膜缘有角膜干细胞存在。

Abstract In order to select stem cells of corneal epithelium which are highly proliferative,and long-live cells,as a source of cell culture,proliferative rates among the cultured limbal,peripheral and central corneal epithelial cells were compared.Methods 5-bromo-2-deoxyuridene(Brdu) immunostaining was used for flow cytometric analysis of proliferative rates of epithelial cells from different areas of cornea of rabbits.Results: the proliferative rates between primary cultured and limbal peripheral corneal cells 48 hours in culture did not appear very different statistically ( P >0 05),but differences of proliferative rates came quite apperent among the limbal,peripheral and central corneal cells soom after the first passage of the cells.The limbal cells were the longest cells in the pass of culture.Conclusion: 1)The proliferative rate of the limbal cells was higher than those of peripheral and central. 2)The slow-cycling cells were supposed to be located in the corneal limbus.Therefore,the stem cells of corneal epithelium are located in the limbus. proliferative rate;corneal epithelial cell;in culture