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为观察离体角膜上皮细胞经传代培养后细胞死亡的规律及确定其性质。本文应用流式细胞仪 ( FACStarplus)检测凋亡细胞的亚 2倍体 DNA以及细胞凋亡的形态学观察 ,对体外培养的每一代兔角膜缘上皮细胞的凋亡情况进行了观察及分析。结果显示 :角膜缘原代上皮细胞被检测出 7.0 0± 2 .2 3 %的凋亡细胞 ;从第 6代开始 ,角膜上皮细胞凋亡数目显著升高 ( P<0 .0 1) ;之后细胞凋亡数目急剧增加 ,第 8～ 9代后细胞出现集体“自杀”现象。透射电镜观察到这些细胞出现了核浓缩、胞膜发泡、凋亡小体等典型细胞凋亡的形态学改变。结论 :离体角膜缘上皮细胞因外部生长环境的改变 ,在培养到第 6代时出现了细胞凋亡现象。
To observe the law of cell death of cultured corneal epithelial cells after subculture and to determine their properties. In this paper, FACStarplus was used to detect the sub-diploid DNA of apoptotic cells and the morphological observation of apoptosis. The apoptosis of limbal epithelial cells of each generation cultured in vitro was observed and analyzed. The results showed that 7.0 ± 2.23% of apoptotic cells were detected in primary limbal epithelial cells. From the sixth passage, the number of apoptotic corneal epithelial cells was significantly increased (P <0.01) The number of apoptotic cells increased sharply. After the 8th to 9th generation, the cells appeared the phenomenon of collective “suicide”. Transmission electron microscopy showed morphological changes in these cells such as nuclear condensation, membrane vesicles, apoptotic bodies and other typical apoptotic cells. CONCLUSION: The changes of limbal epithelial cells in vitro are caused by the change of external environment.