猪视网膜色素上皮细胞的体外培养

目的：将猪视网膜色素上皮(RPE)细胞进行体外培养，为研究RPE细胞的功能及治疗有关眼底疾病提供细胞来源。方法：采用胰酶2次消化法分离猪RPE细胞，15％DMEM培养液培养，每天倒置显微镜观察细胞生长情况，并作流式细胞仪分析培养细胞的细胞周期。以免疫组织化学法鉴定培养细胞的来源。结果：离体培养细胞初期呈圆形，富含黑色素，12～24h贴壁呈圆形、梭形及不规则形。传代后细胞渐趋透明。免疫组化证实RPE细胞均为鼠抗人角蛋白染色阳性。细胞周期分析提示体外培养的RPE细胞保持正常的繁殖能力。结论：培养的细胞可作为体外猪RPE细胞的来源。

Culture of pig retinal pigment epithelial cell in vitro

Objective:In order to provide a large quantity of retinal pigment epithelial (RPE) cells in vitro,we want to establish methods for culture of pig RPE cells.Methods:RPE cells were separated with trypsin twice and were cultured in 15% DMEM medium.The growth of cells was observed under microscope and flow cytometry was used to analyze the cell cycle.The derivation of the cells was proved by immunohistochemical staining with anti-human keratin.Results:The primary cells contained a great amount of pigments in round shape.After 12~24 hours the cells attached took shapes of many kinds.Passages of cultured RPE cells presented transparent shape step by step.Immunohistochmical staining demonstrated that cells were stained by anti-human keratin.The analysis of cell cycle showed the cultured RPE cells keep normal ability of proliferation.Conclusion:The cultured cells may be the foundation of pig RPE cells.