Small-molecule–directed, efficient generation of retinal pigment epithelium from human pluripotent stem cells

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 p...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2015-09, Vol.112 (35), p.10950-10955
Main Authors: 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.
Format: Article
Language:English
Subjects:
Biological Sciences
Cell culture
Cell Differentiation - drug effects
Growth factors
High-Throughput Screening Assays
Humans
Hypoxia
Macular degeneration
Molecules
Pluripotent Stem Cells - cytology
Pluripotent Stem Cells - drug effects
Polymerase Chain Reaction
Retinal Pigment Epithelium - cytology
Stem cells
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Summary: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 onehalf 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.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.1422818112