Reichman Sacha, Goureau Olivier
Institut de la Vision, Institut National de la Santé et de la Recherche Médicale, U968 , 17 Rue Moreau, 75012, Paris, France.
Université Pierre-et-Marie-Curie Paris 6, Sorbonne Universités, UMR S968 , Paris, France.
Methods Mol Biol. 2016;1357:339-51. doi: 10.1007/7651_2014_143.
Human induced pluripotent stem (hiPS) cells could be used as an unlimited source of retinal cells for the treatment of retinal degenerative diseases. Although much progress has been made in the differentiation of pluripotent stem cells towards different retinal lineages, the production of retinal cells from hiPS cells for therapeutic approaches require the development of easy and standardized protocols. In this chapter, we describe a simple and effective protocol for retinal differentiation of hiPS cells bypassing embryoid body formation and the use of exogenous molecules and substrates. In 2 weeks, confluent hiPS cells cultured in pro-neural medium can generate both retinal pigmented epithelial cells and self-forming neural retina-like structures containing retinal progenitor cells. These progenitors can be differentiated into all retinal cell types, including retinal ganglion cells and precursors of photoreceptors, which could find important applications in regenerative medicine. This differentiation system and the resulting hiPS-derived retinal cells will also offer opportunity to study the molecular and cellular mechanisms underlying human retinal development, and the establishment of in vitro models of human retinal degenerative diseases.
人诱导多能干细胞（hiPS细胞）可作为视网膜细胞的无限来源，用于治疗视网膜退行性疾病。尽管在多能干细胞向不同视网膜谱系的分化方面已经取得了很大进展，但从hiPS细胞中产生用于治疗方法的视网膜细胞需要开发简单且标准化的方案。在本章中，我们描述了一种简单有效的hiPS细胞视网膜分化方案，该方案绕过了胚状体的形成以及外源性分子和底物的使用。在两周内，在神经前体细胞培养基中培养的汇合hiPS细胞可以产生视网膜色素上皮细胞和包含视网膜祖细胞的自形成神经视网膜样结构。这些祖细胞可以分化为所有视网膜细胞类型，包括视网膜神经节细胞和光感受器前体，这在再生医学中可能具有重要应用。这种分化系统以及由此产生的hiPS衍生视网膜细胞也将为研究人类视网膜发育的分子和细胞机制以及建立人类视网膜退行性疾病的体外模型提供机会。
