title: Generation of highly enriched populations of optic vesicle-like retinal cells from human pluripotent stem cells.
abstract: The protocol outlined below is used to differentiate human pluripotent stem cells (hPSCs) into retinal cell types through a process that faithfully recapitulates the stepwise progression observed in vivo. From pluripotency, cells are differentiated to a primitive anterior neural fate, followed by progression into two distinct populations of retinal progenitors and forebrain progenitors, each of which can be manually separated and purified. The hPSC-derived retinal progenitors are found to self-organize into three-dimensional optic vesicle-like structures, with each aggregate possessing the ability to differentiate into all major retinal cell types. The ability to faithfully recapitulate the stepwise in vivo development in a three-dimensional cell culture system allows for the study of mechanisms underlying human retinogenesis. Furthermore, this methodology allows for the study of retinal dysfunction and disease modeling using patient-derived cells, as well as high-throughput pharmacological screening and eventually patient-specific therapies.
authors: Department of Biology, Indiana University-Purdue University Indianapolis, Indiana.
authors: Sarah K
authors: SK
authors: Ohlemacher
authors: Department of Biology, Indiana University-Purdue University Indianapolis, Indiana.
authors: Clara L
authors: CL
authors: Iglesias
authors: Department of Biology, Indiana University-Purdue University Indianapolis, Indiana.
authors: Akshayalakshmi
authors: A
authors: Sridhar
authors: Waisman Center, University of Wisconsin-Madison, Wisconsin.|McPherson Eye Research Institute, University of Wisconsin-Madison, Wisconsin.|Department of Ophthalmology and Visual Sciences, University of Wisconsin-Madison, Wisconsin.
authors: David M
authors: DM
authors: Gamm
authors: Department of Biology, Indiana University-Purdue University Indianapolis, Indiana.|Stark Neurosciences Research Institute, Indiana University, Indianapolis, Indiana.|Department of Medical and Molecular Genetics, Indiana University, Indianapolis, Indiana.
authors: Jason S
authors: JS
authors: Meyer
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D019070:Cell Lineage; D049109:Cell Proliferation; D058732:Embryoid Bodies / Q000166:cytology; D006801:Humans; D046569:Neuroepithelial Cells / Q000166:cytology; D009898:Optic Disk / Q000166:cytology*; D039904:Pluripotent Stem Cells / Q000166:cytology*; D012160:Retina / Q000166:cytology*; D055213:Retinal Pigment Epithelium / Q000166:cytology; D018874:Spheroids, Cellular / Q000166:cytology; D013234:Stem Cells / Q000166:cytology
keywords: development; differentiation; human pluripotent stem cells (hPSCs); optic vesicle; retina
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