条件培养液对人视网膜前体细胞分化的诱导

目的:研究各种培养液对人视网膜前体细胞(RPC)的诱导分化作用.方法:分离培养3～5个月人胚胎RPC,分别用胎牛血清(FBS组)、视网膜组织培养上清(RE组)、视网膜色素上皮培养上清(RPE组)诱导分化.采用免疫组化SP法检测细胞诱导前后Nestin、微管相关蛋白-2(MAP-2)、胶质纤维酸性蛋白(GFAP)、视紫红质(Rho-dopsin)及蛋白激酶C(PKC)和钙结合蛋白(Calbindin)的表达情况,流式细胞术检测诱导后MAP-2、GFAP及Rhodopsin阳性细胞数.结果:原代及传代培养的RPC表达Nestin,能分裂增殖,形成子代细胞团.RE组和FBS组细胞诱导分化后形态多样,连接广泛;RPE组细胞呈圆形,不贴壁.诱导分化后RE组细胞MAP-2的表达高于RPE组和FBS组(P＜0.01),而Rhodopsin的表达低于RPE组(P＜0.01);GFAP的表达在RPE组最低,RE组较高,FBS组最高(P＜0.05).结论:不同培养基诱导可使人RPC的分化产生差异.

Differentiation of human retinal progenitor cells induced by different conditional culture medium

Aim: To study the differentiation of human retinal progenitor cells (RPCs) under different culture condition. Methods: Human RPCs isolated from 3 to 5 months-feta were cultured in fetal bovine serum (FBS group), supernatant of cultured retinal explants (RE group), and supernatant of cultured retinal pigment epithelial (RPE) cells (RPE group). The differentiation of RPCs was observed by immunohistochemitry and flow cytometry.Results: The primary and subcultured human RPCs expressed Nestin,and could proliferate and form neurosphere-like cell clusters. Various cells and intercellular connections had been found in RE group and FBS group,but the cells in RPE group were round and floated in the culture medium. The expression of microtubule-associated protein-2 in RE group was significantly higher than those of RPE group and FBS group,the expression of Rhodopsin in RE group was lower than that of RPE group(P<0.01). The expression of glial fibrillary acidic protein in RPE group was the lowest, and the highest in FBS group(P<0.05). Conclusion: The differentiation of human RPCs could be modulated by different culture mediums.