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Triphasic developmentally guided protocol to generate retinal pigment epithelium from induced pluripotent stem cells

Ruchi Sharma, Devika Bose, Jair Montford, Davide Ortolan, Kapil Bharti

2022STAR Protocols39 citationsDOIOpen Access PDF

Abstract

RPE tissues are derived from induced pluripotent stem cells (iPSCs) to model retinal diseases and as a replacement therapy for macular degeneration. Here, we developed a robust and efficient directed differentiation protocol to generate pure RPE cells that form a polarized monolayer. This protocol describes how to set up RPE differentiation and to obtain a pure population that expresses mature RPE markers and forms strong tight junctions. For complete details on the use and execution of this protocol, please refer to Sharma et al., 2019, Sharma et al., 2021 and Miyagishima et al. (2021).

Topics & Concepts

Induced pluripotent stem cellRetinal pigment epitheliumRetinalCell biologyStem cellBiologyPopulationHuman Induced Pluripotent Stem CellsEmbryonic stem cellMedicineGeneticsBiochemistryGeneEnvironmental healthRetinal Development and DisordersCRISPR and Genetic EngineeringPhotoreceptor and optogenetics research