Transcription factor overexpression drives reliable differentiation of retinal pigment epithelium from human induced pluripotent stem cells

Stem Cell Res. 2021 May;53:102368. doi: 10.1016/j.scr.2021.102368. Epub 2021 Apr 27.


Age-related macular degeneration and genetic forms of blindness such as Best Disease and Retinitis Pigmentosa can be caused by degeneration of the Retinal Pigment Epithelium (RPE). RPE generated from patient-derived induced pluripotent stem cells (iPSCs) is valuable for both the study of disease mechanisms and development of therapeutic strategies. However, protocols to produce iPSC-derived RPE in vitro are often inefficient, labor-intensive, low-throughput, and highly variable between cell lines and within batches. Here, we report a robust, scalable method to generate iPSC-RPE using doxycycline-inducible expression of eye field transcription factors OTX2, PAX6 and MITF paired with RPE-permissive culture media. Doxycycline addition induces exogenous expression of these transcription factors in Best Disease patient- and wildtype iPSCs to efficiently produce monolayers of RPE with characteristic morphology and gene expression. Further, these RPE monolayers display functionality features including light absorption via pigmentation, polarity-driven fluid transport, and phagocytosis. With this method, we achieve a highly efficient and easily scalable differentiation without the need for mechanical isolation or enrichment methods, generating RPE cultures applicable for in vitro studies.

Keywords: Cell Differentiation; Induced Pluripotent Stem Cells; Retinal Diseases; Retinal Pigment Epithelium; Transcription Factors; iPSC.

MeSH terms

  • Cell Differentiation
  • Cell Line
  • Humans
  • Induced Pluripotent Stem Cells*
  • Retinal Pigment Epithelium
  • Transcription Factors / genetics


  • Transcription Factors