培养的角膜缘上皮细胞增殖能力的动态研究

A dynamic study on proliferative ability of cultivated limbal epithelial cells

目的 探讨组织块培养角膜缘上皮细胞在不同时间以及第 2、3代培养的角膜缘上皮细胞早期 (6～ 8d)的增殖能力。方法 应用组织块培养法培养角膜缘上皮细胞 ,在d8、12、16、2 0、2 4用流式细胞仪检测其细胞周期 ,同时对组织块培养 16d的角膜缘上皮细胞进行传代 ,用同样方法检测第 2、3代细胞出现汇合时 (6～ 8d)的细胞周期。结果 组织块培养角膜缘上皮细胞增殖指数 (ProliferativeIndex ,PI)在高水平上呈下降趋势 (分别为 31 2 %、2 0 1%、17 6 %、15 6 %、13 4 % ) ;2、3代培养细胞早期的增殖指数较高 (分别为 30 3%、2 6 % )。结论 组织块培养的角膜缘上皮细胞增殖能力在 8～ 2 4d内在高水平上呈下降趋势 ,第 2、3代培养的细胞在培养早期的增殖能力也较高 。

Objective To investigate proliferative ability of cultivated limbal epithelial cells(LEC) by tissue inoculation at the different time and of cultivated limbal epithelial cells of the second and third passages in the early period(6～8d). Methods Limbal epithelial cells were cultivated by tissue inoculation method, their cell cycle was checked with lapse cytometry on the 8 th,12 th,16 th,20 th and 24th day, and meanwhile cultivated limbal epithelial cells by tissue inoculation were passaged on the 16 th day, the cell cycle of the second and the third passages was checked by the same method. Results Proliferative index (PI) of cultivated LEC by tissue inoculation on the 8 th,12 th,16 th,20 th and 24 th days were higher and showed the downtrend(31 2%,20 1%,17 6%,15 6% and 13 4% respectively);PI of the second and the third passages were also higher(30 3% and 26% respectively). Conclusion Proliferative ability of cultivated limbal epithelial cells by tissue inoculation within 8～24 d is at the higher level and shows the downtrend, proliferative ability of the second and the third passages in the early period(6～8 d) is also higher, which can be used for transplantation.