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 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.
Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287;
Julien
J
Maruotti
Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287;
Srinivas R
SR
Sripathi
Unit on Ocular and Stem Cell Translational Research, National Eye Institute, National Institutes of Health, Bethesda, MD 20892;
Kapil
K
Bharti
Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287;
John
J
Fuller
Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287;
Karl J
KJ
Wahlin
Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287;
Vinod
V
Ranganathan
Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287;
Valentin M
VM
Sluch
Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287;
Cynthia A
CA
Berlinicke
Unit on Ocular and Stem Cell Translational Research, National Eye Institute, National Institutes of Health, Bethesda, MD 20892;
Janine
J
Davis
Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287;
Catherine
C
Kim
Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287;
Lijun
L
Zhao
Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287;
Jun
J
Wan
Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287;
Jiang
J
Qian
Stem Cell Core Facility, Columbia University, New York, NY 10032;
Barbara
B
Corneo
Neural Stem Cell Institute, Rensselaer, NY 12144;
Sally
S
Temple
Department of Medicine, Stanford University School of Medicine, Stanford, CA 94305; Department of Biochemistry, Stanford University School of Medicine, Stanford, CA 94305;
Ramin
R
Dubey
Department of Pharmacology and Pharmaceutical Sciences, University of Southern California, Los Angeles, CA 90089;
Bogdan Z
BZ
Olenyuk
Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287;
Imran
I
Bhutto
Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287;
Gerard A
GA
Lutty
Wilmer Eye Institute, Johns Hopkins University School of Medicine, Baltimore, MD 21287; Department of Molecular Biology and Genetics, Johns Hopkins University, Baltimore, MD 21287; Department of Neuroscience, Johns Hopkins University, Baltimore, MD 21287; Institute of Genetic Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21287 dzack@jhmi.edu.
Donald J
DJ
Zack
D002454:Cell Differentiation / Q000187:drug effects*; D057166:High-Throughput Screening Assays; D006801:Humans; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects*; D016133:Polymerase Chain Reaction; D055213:Retinal Pigment Epithelium / Q000166:cytology*
age-related macular degeneration; differentiation; high-throughput screening; pluripotent stem cells; retinal pigment epithelium