小分子定向、高效地从人多能干细胞生成视网膜色素上皮。
Small-molecule-directed, efficient generation of retinal pigment epithelium from human pluripotent stem cells.
作者信息
Maruotti Julien, Sripathi Srinivas R, Bharti Kapil, Fuller John, Wahlin Karl J, Ranganathan Vinod, Sluch Valentin M, Berlinicke Cynthia A, Davis Janine, Kim Catherine, Zhao Lijun, Wan Jun, Qian Jiang, Corneo Barbara, Temple Sally, Dubey Ramin, Olenyuk Bogdan Z, Bhutto Imran, Lutty Gerard A, Zack Donald J
机构信息
Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287;
Unit on Ocular and Stem Cell Translational Research, National Eye Institute, National Institutes of Health, Bethesda, MD 20892;
出版信息
Proc Natl Acad Sci U S A. 2015 Sep 1;112(35):10950-5. doi: 10.1073/pnas.1422818112. Epub 2015 Aug 12.
Age-related macular degeneration (AMD) is associated with dysfunction and death of retinal pigment epithelial (RPE) cells. Cell-based approaches using RPE-like cells derived from human pluripotent stem cells (hPSCs) are being developed for AMD treatment. However, most efficient RPE differentiation protocols rely on complex, stepwise treatments and addition of growth factors, whereas small-molecule-only approaches developed to date display reduced yields. To identify new compounds that promote RPE differentiation, we developed and performed a high-throughput quantitative PCR screen complemented by a novel orthogonal human induced pluripotent stem cell (hiPSC)-based RPE reporter assay. Chetomin, an inhibitor of hypoxia-inducible factors, was found to strongly increase RPE differentiation; combination with nicotinamide resulted in conversion of over one-half of the differentiating cells into RPE. Single passage of the whole culture yielded a highly pure hPSC-RPE cell population that displayed many of the morphological, molecular, and functional characteristics of native RPE.
年龄相关性黄斑变性（AMD）与视网膜色素上皮（RPE）细胞的功能障碍和死亡有关。目前正在开发使用源自人类多能干细胞（hPSC）的类RPE细胞的基于细胞的方法来治疗AMD。然而，最有效的RPE分化方案依赖于复杂的逐步处理和添加生长因子，而迄今为止开发的仅使用小分子的方法产量较低。为了鉴定促进RPE分化的新化合物，我们开发并进行了高通量定量PCR筛选，并辅以基于新型正交人诱导多能干细胞（hiPSC）的RPE报告基因检测。发现缺氧诱导因子抑制剂切托明可强烈增加RPE分化；与烟酰胺联合使用可使超过一半的分化细胞转化为RPE。对整个培养物进行单次传代可产生高度纯化的hPSC-RPE细胞群体，该群体表现出许多天然RPE的形态、分子和功能特征。
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