bFGF诱导人胚视网膜色素上皮细胞转分化的研究

目的 探讨bFGF体外诱导早期人胚视网膜色素上皮（RPE）细胞转分化现象及其分子机制,为视网膜发育的进一步基础研究和临床RPE细胞移植奠定基础.方法 取4～6代人胚（6 w～10 w）RPE细胞,bFGF（20 ng/ml）诱导培养（4 d～6 d）后观察RPE细胞形态学的改变,用免疫细胞化学作转分化RPE细胞鉴定.RT-PCR分析转分化RPE细胞小眼畸形基因（MITF）的表达.结果 bFGF诱导培养后的人胚RPE细胞呈现类神经样细胞表型,不仅表达自身上皮细胞标记物角蛋白（Keratin）,同时还表达多种神经视网膜上皮细胞标记物（MAP2、GS、GFAP、Peripherin、Opsin,但不表达NCAM）;用RT-PCR方法没有检测到转分化RPE细胞MITF-A mRNA表达.结论 早期人胚RPE细胞的发育具有一定的可塑性,bFGF可以诱导早期人胚RPE细胞表达多种神经视网膜上皮细胞标记物,可能是通过抑制MITF-A的分子信号通路.

bFGF Induced Transdifferentiation of Human Embryonic Retinal Pigment Epithelium Cells

Objective To study the bFGF induced transdifferentiation of human embryonic retinal pigment epithelium cells(hRPE) cells,with the intention to expand the possibility of the therapeutic RPE cells transplantation.Methods Administrating optimized dose of bFGF to the RPE cells for the transdifferentiation study,we studied the neuronal retina markers after the transdifferentiation of RPE cells with immunohistochemistry staining.Using a semi-quantitative RT-PCR method,we analyzed the mRNA expression of MITF-A in the differentiation RPE cells.Results RPE cells co-expressed both the RPE cells marker cytokeratin and the neuronal retina markers(MAP2,GS,GFAP,Peripherin,Opsin),when stimulated by bFGF.We did not detect the expression of NCAM in the transdifferentiation RPEs using immunohistochemistry.The expression of MITF-A mRNA was not detected in the bFGF treated RPE cells when analyzed by RT-PCR.Conclusion Expressions of both neuronal retina and RPE markers in cultured embryonic hRPE cells induced by bFGF might be through the inhibition of MITF-A pathway.