摘要
目的 观察层粘连蛋白 (LN)对离体角膜上皮细胞存亡的影响 ,以寻找预防角膜上皮细胞凋亡的途径。方法 应用流式细胞仪 (FACStarplus)检测凋亡细胞的亚 2倍体DNA的方法 ,对基础培养的、与LN共育的以及阻断LN受体后的兔角膜上皮细胞的凋亡情况进行了对比观察及分析。结果 基础培养的第 6代角膜上皮细胞出现明显的细胞凋亡 ;与LN共育的角膜上皮细胞凋亡不明显 ;阻断LN受体后 ,角膜上皮细胞出现显著的凋亡现象。结论 LN对离体培养的角膜上皮细胞的凋亡有保护作用。
ObjectiveTo prevent apoptosis of corneal epithelial cells cultured in vitro,the effects of laminin upon cell survival was studied.MethodsCorneal epithelial cells from New Zealand white rabbits were cultured respectively in basic culture medium (DMEM Ham’s F 12 medium 1∶1),in basic culture medium in which laminin was added and in basic culture medium in which antibody against laminin was added.The cells were passaged every?48 h.The cells form passages 1 to 6 were fixed with 70% ethanol after cultured for 48?h.After staining with propidium iodide(PI),apoptosis was measured by the presence of degraded DNA(<?2?n DNA) in a flow cytometric assay.ResultsFlow cytometric analysis of propidium iodide stained cells indicated that 7 00%±2 23% of the primary cells contained <?2?n DNA while cultured in basic culture medium.Sixth passage cells grown in basic culture medium showed significantly increased amounts of degraded DNA (18 74%±3 93%, P <0 01).A significantly higher percentage of cells survived in the 6th passage when cells were cultured in the presence of laminin.In contrast,an increased amount of apoptosis(30 80%±7 90%)was found when cells were cultured in the presence of with anti laminin antibody.ConclusionLaminin can prevent apoptosis of corneal epithelial cells cultured in vitro.
出处
《眼科研究》
CAS
CSCD
2000年第2期112-113,共2页
Chinese Ophthalmic Research
基金
国家自然科学基金!资助 (基金编号 :39870 855)
广东省科学基金!资助 (基金编号 :930 4 4 3)
