Defined Medium Conditions for the Induction and Expansion of Human Pluripotent Stem Cell-Derived Retinal Pigment Epithelium

Grace E. Lidgerwood, Shiang Y. Lim, Duncan E. Crombie, Ray Ali, Katherine P. Gill, Damián Hernández, Josh Kie, Alison Conquest, Hayley S. Waugh, Raymond C B Wong, Helena H. Liang, Alex W. Hewitt, Kathryn C. Davidson, Alice Pébay

Research output: Contribution to journalReview ArticleResearchpeer-review

13 Citations (Scopus)

Abstract

We demonstrate that a combination of Noggin, Dickkopf-1, Insulin Growth Factor 1 and basic Fibroblast Growth Factor, promotes the differentiation of human pluripotent stem cells into retinal pigment epithelium (RPE) cells. We describe an efficient one-step approach that allows the generation of RPE cells from both human embryonic stem cells and human induced pluripotent stem cells within 40–60 days without the need for manual excision, floating aggregates or imbedded cysts. Compared to methods that rely on spontaneous differentiation, our protocol results in faster differentiation into RPE cells. This pro-retinal culture medium promotes the growth of functional RPE cells that exhibit key characteristics of the RPE including pigmentation, polygonal morphology, expression of mature RPE markers, electrophysiological membrane potential and the ability to phagocytose photoreceptor outer segments. This protocol can be adapted for feeder, feeder-free and serum-free conditions. This method thereby provides a rapid and simplified production of RPE cells for downstream applications such as disease modelling and drug screening.

Original languageEnglish
Pages (from-to)179-188
Number of pages10
JournalStem Cell Reviews and Reports
Volume12
Issue number2
DOIs
Publication statusPublished - 1 Apr 2016

Keywords

  • Differentiation
  • Electrophysiology
  • Human embryonic stem cells
  • Human induced pluripotent stem cells
  • Retinal pigment epithelium
  • RNA sequencing

Cite this

Lidgerwood, G. E., Lim, S. Y., Crombie, D. E., Ali, R., Gill, K. P., Hernández, D., Kie, J., Conquest, A., Waugh, H. S., Wong, R. C. B., Liang, H. H., Hewitt, A. W., Davidson, K. C., & Pébay, A. (2016). Defined Medium Conditions for the Induction and Expansion of Human Pluripotent Stem Cell-Derived Retinal Pigment Epithelium. Stem Cell Reviews and Reports, 12(2), 179-188. https://doi.org/10.1007/s12015-015-9636-2