Department of Biology, Indiana University-Purdue University Indianapolis, Indianapolis, IN, United States.
Department of Medical and Molecular Genetics, Indiana University School of Medicine, Indianapolis, IN, United States; Department of Ophthalmology, Glick Eye Institute, Indiana University School of Medicine, Indianapolis, IN, United States; Stark Neurosciences Research Institute, Indiana University School of Medicine, Indianapolis, IN, United States.
Methods Cell Biol. 2020;159:279-302. doi: 10.1016/bs.mcb.2020.02.005. Epub 2020 Mar 11.
Human pluripotent stem cells (hPSCs) possess the remarkable ability to differentiate into any cell type of the body, including those of the retina. Through the differentiation of these cells as retinal organoids, it is now possible to model the spatial and temporal development of the human retina using hPSCs, in which retinal progenitor cells produce the entire repertoire of retinal cells, first differentiating into retinal ganglion cells and ending with mature photoreceptors, bipolar cells, and Müller glia. Importantly, retinal organoids self-assemble into laminated structures that recapitulate the layering of the human retina with a retinal ganglion cell layer lining the inner layer and a distinctly separate photoreceptor layer occupying the outer layers. This organoid technology has provided access to human tissue for developmental and disease modeling, as well as translational applications such as high throughput drug screening and cell replacement therapies. However, the differentiation of retinal organoids does require some expertise and multiple strategies produce inconsistent results. Here, we describe in detail a well-established and relatively simple method for the generation of retinal organoids.
人类多能干细胞（hPSCs）具有分化为身体任何细胞类型的显著能力，包括视网膜细胞。通过将这些细胞分化为视网膜类器官，可以使用 hPSCs 对人类视网膜的时空发育进行建模，其中视网膜祖细胞产生整个视网膜细胞库，首先分化为视网膜神经节细胞，最后分化为成熟的光感受器、双极细胞和 Muller 胶质细胞。重要的是，视网膜类器官会自动组装成层状结构，重现人类视网膜的分层，内层是视网膜神经节细胞层，外层是明显分离的光感受器层。这项类器官技术为发育和疾病建模以及转化应用（如高通量药物筛选和细胞替代疗法）提供了人类组织的获取途径。然而，视网膜类器官的分化确实需要一些专业知识，并且多种策略会产生不一致的结果。在这里，我们详细描述了一种成熟且相对简单的生成视网膜类器官的方法。
