兔视网膜色素上皮细胞培养和鉴定

目的：进行兔视网膜色素上皮细胞(RPE)细胞的体外培养，探讨RPE培养与鉴定方法，为研究其生理功能、生化特征和病理过程提供细胞模型。方法：用改良的眼杯固定法，将其固定在眼球托上，用胰酶直接消化视网膜色素上皮面，收取RPE细胞。作细胞形态学、免疫组织化学、免疫荧光鉴定。结果：RPE细胞培养早期生长活跃、胞核透明、胞浆含丰富黑色素颗粒，多巴氧化酶呈阳性反应．免疫组织化学染色和免疫荧光染色提示角蛋白表达强阳性。结论：培养的大量细胞可用于RPE的体外实验研究，多巴染色是鉴定RPE细胞一种较好的方法。

In vitro culture and identification of retinal pigment epithelium cells in rabbits

Objective:To establish the cultivating method of retinal pigment epithelium cells( RPE)in rabbits.Methods:A paralimbal incision was made and extended circumferentially to separate the anterior segment on a sterile filter paper.Then the posterior eye cup was fixed in a rubber plug.The single cell was collected directly by trypsinization and cultured in 20% DMEM/F12 medium. The RPE cells were observed under microscope and identified by DOPA staining and immunohistochemistry.Results:RPE cells grew in a well-distributed way rather than in colony in experimental conditions.The primary cells contained a great amount of melanin in round shape.DOPA staining showed positive and immunohistochemical staining demonstrated that cells were stained by anti-human keratin.Conclusion:We can produce a large quantity of RPE cells for in vitro experiments and DOPA staining proves effective in identification of RPE cells.