兔虹膜和视网膜色素上皮细胞的体外培养比较

目的 观察比较有色素兔虹膜色素上皮(IPE)及视网膜色素上皮(RPE)细胞的体外生长状况。方法 采用酶消化法及酶辅助机械分离法分别分离IPE和RPE细胞。观察培养的两种细胞的生长状况，并对其进行免疫组化鉴定。结果 原代IPE及RPE细胞大多呈圆形或六边形，细胞内充满了黑色素颗粒。随着传代的增加，呈梭形或纤维细胞样生长的细胞增多，细胞中的色素颗粒也逐渐减少。细胞角蛋白免疫组化染色显示IPE及RPE细胞绝大多数呈棕黄色阳性反应。Desmin兔疫组化染色显示IPE细胞中阳性细胞约占5％。结论 分离培养的IPE和RPE细胞纯度很高。IPE细胞中绝大多数为后层IPE细胞。IPE和RPE细胞的形态及体外生长特点类似。

Culture of IPE and RPE cells from pigmented rabbits

Objective To isolate and culture IPE and RPE cells of pigmented rabbits and observe and compare their growth characteristics in vitro. Methods IPE and RPE cells were isolated by enzymatic digestion method and enzyme-assisted microdissection method respectively. Both IPE and RPE cells were cultured in DMEM/F12 medium supplemented with 20% newborn calf serum. The cells were identified as epithelia cell by immunocytochemical method. Results There were a great number of round, pigmented IPE and RPE cells and some cell clusters. IPE and RPE cells began to attach to culture dishes in more than 12 hours. They spread into polygonal,round,spindle-shaped,etc. These cells had round nuclear and dense pigmentation. They grew very rapidly at 4 ' to 6th day. When the cells were passaged into subcultures, the pigment granules were gradually lighter and the spindle-shaped or fiber-like cells became more. Both IPE and RPE cells stained positive for cytokeratin. About 5% IPE cells stained positive for desmin. Conclusions The cultured IPE and RPE cells are pure. Their growth characteristics is similar. Most of IPE cells are from posterior layer.