人胚胎视网膜色素上皮细胞的体外培养和吞噬作用

In vitro culture and phagocytosis of human embryonic retinal pigment epithelium

目的探讨人胚胎视网膜色素上皮(RPE)细胞分离培养的特性及其吞噬作用。方法取发育至13～15周的新鲜人胚胎眼球,显微解剖后采用机械胰酶消化法分离培养RPE细胞,应用免疫荧光法观察人胚胎RPE细胞吞噬视网膜光感受细胞外节膜盘。结果人胚胎发育至13～15周后,RPE细胞可应用标准的培养方法进行体外培养和传代。根据测定的人胚胎RPE细胞生长曲线,取2～6代细胞用于实验培养的人胚胎RPE细胞具备吞噬功能。结论改进了人胚胎RPE细胞的培养方法,证明人胚胎RPE细胞在体外具有吞噬能力。

Objective To analyze the culturing characteristics and phagocytosis of the retinal pigment epithelium （RPE） from human embryo, and to provide the cellular basis for the further research of RPE related diseases. Methods Fresh human embryonic eyeballs （13-15 weeks） were collected, and RPE cells were separated by mechanical and trypsin digestion after microdissection. Human embryonic RPE phagocytosis of retinal outer segments was investigated by using immunofluorescence. Results After the human embryo had developed for 13-15 weeks, the RPE cells could be cultured in vitro according to standard culturing methods. Based on the growth curve, the RPE cells in generations 2-6 were collected for essays and showed the ability of phagocytosis afterwards. Conclusion The method of culturing human embryonic RPE cells is improved, and phagocytosis ability of human embryonic RPE cells is also proved in vitro according to this study.