UCL Discovery
UCL home » Library Services » Electronic resources » UCL Discovery

Directing Differentiation of Pluripotent Stem Cells Toward Retinal Pigment Epithelium Lineage

Choudhary, P; Booth, H; Gutteridge, A; Surmacz, B; Louca, I; Steer, J; Kerby, J; (2016) Directing Differentiation of Pluripotent Stem Cells Toward Retinal Pigment Epithelium Lineage. Stem Cells Translational Medecine , 6 (2) pp. 490-501. 10.5966/sctm.2016-0088. Green open access

[thumbnail of Choudhary_et_al-2017-STEM_CELLS_Translational_Medicine.pdf]
Preview
Text
Choudhary_et_al-2017-STEM_CELLS_Translational_Medicine.pdf - Published Version

Download (5MB) | Preview

Abstract

Development of efficient and reproducible conditions for directed differentiation of pluripotent stem cells into specific cell types is important not only to understand early human development but also to enable more practical applications, such as in vitro disease modeling, drug discovery, and cell therapies. The differentiation of stem cells to retinal pigment epithelium (RPE) in particular holds promise as a source of cells for therapeutic replacement in age-related macular degeneration. Here we show development of an efficient method for deriving homogeneous RPE populations in a period of 45 days using an adherent, monolayer system and defined xeno-free media and matrices. The method utilizes sequential inhibition and activation of the Activin and bone morphogenetic protein signaling pathways and can be applied to both human embryonic stem cells and induced pluripotent stem cells as the starting population. In addition, we use whole genome transcript analysis to characterize cells at different stages of differentiation that provides further understanding of the developmental dynamics and fate specification of RPE. We show that with the described method, RPE develop through stages consistent with their formation during embryonic development. This characterization- together with the absence of steps involving embryoid bodies, three-dimensional culture, or manual dissections, which are common features of other protocols-makes this process very attractive for use in research as well as for clinical applications. SIGNIFICANCE: This report describes a novel method of directed differentiation to generate retinal pigment epithelium (RPE) cells from pluripotent stem cells. The employed method is based on adherent monolayer culture using xeno-free conditions and manipulation of the Activin and bone morphogenetic protein signaling pathway using small molecules and recombinant proteins. Whole genome microarray analysis was performed to characterize the differentiation process and understand the developmental path of RPE generation in vitro. This method can be applied for generation of RPE for research as well as for clinical applications.

Type: Article
Title: Directing Differentiation of Pluripotent Stem Cells Toward Retinal Pigment Epithelium Lineage
Location: United States
Open access status: An open access version is available from UCL Discovery
DOI: 10.5966/sctm.2016-0088
Publisher version: http://dx.doi.org/10.5966/sctm.2016-0088
Language: English
Additional information: © AlphaMed Press. This is an open access article under the terms of the CreativeCommons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Keywords: Activin, Bone morphogenetic protein, Directed differentiation, Retinal pigment epithelium, Stem cells
UCL classification: UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Queen Square Institute of Neurology
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Brain Sciences > UCL Queen Square Institute of Neurology > Department of Neuromuscular Diseases
URI: https://discovery.ucl.ac.uk/id/eprint/1514457
Downloads since deposit
112Downloads
Download activity - last month
Download activity - last 12 months
Downloads by country - last 12 months

Archive Staff Only

View Item View Item