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Defined medium conditions for the induction and expansion of human pluripotent stem cell-derived retinal pigment epithelium

Citation

Lidgerwood, GE and Lim, SY and Crombie, DE and Ali, R and Gill, KP and Hernandez, D and Kie, J and Conquest, A and Waugh, HS and Wong, RCB and Liang, HH and Hewitt, AW and Davidson, KC and Pebay, A, Defined medium conditions for the induction and expansion of human pluripotent stem cell-derived retinal pigment epithelium, Stem Cell Reviews and Reports, 12, (2) pp. 179-188. ISSN 1550-8943 (2016) [Refereed Article]

Copyright Statement

Copyright 2015 Springer Science+Business Media New York

DOI: doi:10.1007/s12015-015-9636-2

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.

Item Details

Item Type:Refereed Article
Keywords:differentiation, electrophysiology, human embryonic stem cells, human induced pluripotent stem cells, RNA sequencing, retinal pigment epithelium
Research Division:Medical and Health Sciences
Research Group:Ophthalmology and Optometry
Research Field:Ophthalmology
Objective Division:Health
Objective Group:Clinical Health (Organs, Diseases and Abnormal Conditions)
Objective Field:Hearing, Vision, Speech and Their Disorders
Author:Ali, R (Dr Ray Ali)
Author:Hewitt, AW (Dr Alex Hewitt)
ID Code:107312
Year Published:2016 (online first 2015)
Web of Science® Times Cited:2
Deposited By:Menzies Institute for Medical Research
Deposited On:2016-03-10
Last Modified:2017-11-07
Downloads:0

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