TY - JOUR
T1 - Small-molecule-directed, efficient generation of retinal pigment epithelium from human pluripotent stem cells
AU - Maruotti, Julien
AU - Sripathi, Srinivas R.
AU - Bharti, Kapil
AU - Fuller, John
AU - Wahlin, Karl J.
AU - Ranganathan, Vinod
AU - Sluch, Valentin M.
AU - Berlinicke, Cynthia A.
AU - Davis, Janine
AU - Kim, Catherine
AU - Zhao, Lijun
AU - Wan, Jun
AU - Qian, Jiang
AU - Corneo, Barbara
AU - Temple, Sally
AU - Dubey, Ramin
AU - Olenyuk, Bogdan Z.
AU - Bhutto, Imran
AU - Lutty, Gerard A.
AU - Zack, Donald J.
N1 - Publisher Copyright:
© 2015, National Academy of Sciences. All rights reserved.
PY - 2015/9/1
Y1 - 2015/9/1
N2 - 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 one-half 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.
AB - 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 one-half 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.
KW - Age-related macular degeneration
KW - Differentiation
KW - High-throughput screening
KW - Pluripotent stem cells
KW - Retinal pigment epithelium
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U2 - 10.1073/pnas.1422818112
DO - 10.1073/pnas.1422818112
M3 - Article
C2 - 26269569
AN - SCOPUS:84941209330
SN - 0027-8424
VL - 112
SP - 10950
EP - 10955
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 35
ER -