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

Proc Natl Acad Sci U S A. 2015 Sep 1;112(35):10950-5. doi: 10.1073/pnas.1422818112. Epub 2015 Aug 12.


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.

Keywords: age-related macular degeneration; differentiation; high-throughput screening; pluripotent stem cells; retinal pigment epithelium.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Differentiation / drug effects*
  • High-Throughput Screening Assays
  • Humans
  • Pluripotent Stem Cells / cytology
  • Pluripotent Stem Cells / drug effects*
  • Polymerase Chain Reaction
  • Retinal Pigment Epithelium / cytology*