Parvini Maryam, Parivar Kazem, Safari Fatemeh, Tondar Mahdi
Department of Stem Cells and Developmental Biology at Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran,
In Vitro Cell Dev Biol Anim. 2015 Mar;51(3):310-8. doi: 10.1007/s11626-014-9835-1. Epub 2014 Nov 27.
Despite the enormous progress in studying retinal cell differentiation from human embryonic stem cells (hESCs), none of the reported protocols have produced a cost-effective eye field cells with the capability to further differentiate into retinal derivatives. In this study, by drawing chemicals on our four-step differentiation strategy, we demonstrated the ability of hESCs in assembling such qualifications to follow human retinogenesis in a serum- and feeder-free adherent condition. Two-dimensional (2D) populations of eye field cells arose within early forebrain progeny upon hESCs differentiation. Gene expression analysis showed that the treatment of hESCs with a combination of selected small molecules (SMs) gave rise to the higher expressions of eye field-specific genes, PAX6, RX, and SIX3. Thereafter, a subset of cells gained the transient features of advancing retinal differentiation, including optic vesicle (OV)-like structures, which expressed MITF and CHX10 in a manner imitated in vivo human retinal development. The competency of derived cells in differentiation to retinal derivatives was further investigated. The gene analysis of the cells showed more propensity for generating retinal pigment epithelial (RPE) than neural retina (NR). The generation of OV-like structures in 2D cultures can shed light on molecular events governing retinal specification. It can also facilitate the study of human retinal development.
尽管在研究人胚胎干细胞（hESCs）向视网膜细胞分化方面取得了巨大进展，但所报道的方案均未产生具有进一步分化为视网膜衍生物能力的具有成本效益的眼场细胞。在本研究中，通过借鉴我们的四步分化策略中的化学物质，我们证明了hESCs在无血清和无饲养层的贴壁条件下具备遵循人类视网膜发生过程的能力。hESCs分化后，在前脑早期后代中出现了二维（2D）眼场细胞群体。基因表达分析表明，用选定的小分子（SMs）组合处理hESCs会导致眼场特异性基因PAX6、RX和SIX3的更高表达。此后，一部分细胞获得了推进视网膜分化的短暂特征，包括类似视泡（OV）的结构，这些结构以模仿体内人类视网膜发育的方式表达MITF和CHX10。进一步研究了衍生细胞分化为视网膜衍生物的能力。对这些细胞的基因分析表明，它们生成视网膜色素上皮（RPE）的倾向大于神经视网膜（NR）。二维培养中类似视泡结构的产生可以揭示控制视网膜特化的分子事件。它还可以促进对人类视网膜发育的研究。
