Department of Stem Cells and Developmental Biology, The Academic Center for Education, Culture and Research-ACECR, Tehran, Iran.
Stem Cells Dev. 2012 Aug 10;21(12):2262-72. doi: 10.1089/scd.2011.0599. Epub 2012 Feb 3.
We describe a new, efficient protocol that involves the serial addition of noggin, basic fibroblast growth factor (bFGF), retinoic acid, and sonic hedgehog (Shh) for the differentiation of human induced pluripotent stem cells (hiPSC) to retinal pigmented epithelium (RPE) in a serum- and feeder-free adherent condition. hiPSC-RPE cells exhibited RPE morphology and specific molecular markers. Additionally, several hiPSC lines were generated from retinal-specific patients with Leber's congenital amaurosis, Usher syndrome, two patients with retinitis pigmentosa, and a patient with Leber's hereditary optic neuropathy. The RPE cells generated from these disease-specific hiPSCs expressed specific markers by the same RPE lineage-directed differentiation protocol. These findings indicate a new short-term, simple, and efficient protocol for differentiation of hiPSCs to RPE cells. Such specific retinal disease-specific hiPSCs offer an unprecedented opportunity to recapitulate normal and pathologic formation of human retinal cells in vitro, thereby enabling pharmaceutical screening, and potentially autologous cell replacement therapies for retinal diseases.
我们描述了一种新的、高效的方案，涉及到在无血清和无饲养层的贴壁条件下，将 noggin、碱性成纤维细胞生长因子（bFGF）、维甲酸和 sonic hedgehog（Shh）依次添加到诱导多能干细胞（hiPSC）中，使其分化为视网膜色素上皮（RPE）。 hiPSC-RPE 细胞表现出 RPE 形态和特定的分子标记。此外，我们还从 Leber 先天性黑蒙、Usher 综合征、两名视网膜色素变性患者和一名 Leber 遗传性视神经病变患者的视网膜特异性患者中生成了多个 hiPSC 系。通过相同的 RPE 谱系定向分化方案，这些疾病特异性 hiPSC 产生的 RPE 细胞表达特定的标记。这些发现表明，这是一种新的、短期的、简单而有效的 hiPSC 向 RPE 细胞分化的方案。这些特定的视网膜疾病特异性 hiPSC 为在体外重现人类视网膜细胞的正常和病理形成提供了前所未有的机会，从而实现了药物筛选，并可能为视网膜疾病提供自体细胞替代疗法。
