April 2010
Volume 51, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2010
Isolation, Differentiation and Developmental Modeling of Early Retinal Progenitor Cell Populations From Human Pluripotent Stem Cells
J. S. Meyer; E. E. Capowski; K. A. Wallace; A. D. Verhoeven; A. V. Sloman; J. M. Martin; L. S. Wright; R. Stewart; J. A. Thomson; D. M. Gamm
Author Affiliations & Notes
  • J. S. Meyer
    Waisman Center,
    University of Wisconsin, Madison, Wisconsin
  • E. E. Capowski
    Waisman Center,
    University of Wisconsin, Madison, Wisconsin
  • K. A. Wallace
    Waisman Center,
    University of Wisconsin, Madison, Wisconsin
  • A. D. Verhoeven
    Waisman Center,
    University of Wisconsin, Madison, Wisconsin
  • A. V. Sloman
    Waisman Center,
    University of Wisconsin, Madison, Wisconsin
  • J. M. Martin
    Waisman Center,
    University of Wisconsin, Madison, Wisconsin
  • L. S. Wright
    Waisman Center,
    University of Wisconsin, Madison, Wisconsin
  • R. Stewart
    Morgridge Institute,
    University of Wisconsin, Madison, Wisconsin
  • J. A. Thomson
    Morgridge Institute,
    University of Wisconsin, Madison, Wisconsin
  • D. M. Gamm
    Waisman Center,
    Ophthalmology and Visual Sciences, Eye Research Institute,
    University of Wisconsin, Madison, Wisconsin
  • Footnotes
    Commercial Relationships  J.S. Meyer, None; E.E. Capowski, None; K.A. Wallace, None; A.D. Verhoeven, None; A.V. Sloman, None; J.M. Martin, None; L.S. Wright, None; R. Stewart, None; J.A. Thomson, None; D.M. Gamm, None.
  • Footnotes
    Support  Foundation Fighting Blindness, Lincy Foundation, Research to Prevent Blindness McCormick Scholar Award, Walsh Foundation, Retina Research Foundation and Heckrodt Foundation, and NICHD P30 HD03352
Investigative Ophthalmology & Visual Science April 2010, Vol.51, 1230. doi:
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      J. S. Meyer, E. E. Capowski, K. A. Wallace, A. D. Verhoeven, A. V. Sloman, J. M. Martin, L. S. Wright, R. Stewart, J. A. Thomson, D. M. Gamm; Isolation, Differentiation and Developmental Modeling of Early Retinal Progenitor Cell Populations From Human Pluripotent Stem Cells. Invest. Ophthalmol. Vis. Sci. 2010;51(13):1230.

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Abstract

Purpose: : Established methods for deriving early retinal progenitors from human pluripotent stem cells (hPSCs) typically yield a heterogeneous population of cells, which complicates studies of retinal development. Therefore, we sought to develop a simple method of isolating a highly enriched population of optic vesicle (OV) stage, multipotent retinal progenitor cells from human ES and iPS cells.

Methods: : hPSCs were differentiated toward a retinal lineage using a previously described protocol. Highly enriched populations of OV stage retinal progenitors were manually separated from forebrain progenitor populations and allowed to differentiate for up to 120 days. Differences in gene expression between retinal and forebrain progenitor populations were determined via PCR and microarray analyses. Differentiating OV populations were then examined by qPCR and ICC to ascertain their ability to generate retinal cell types. Subsequently, selected inducing factors were added for discrete periods to study effects on cell fate choice.

Results: : Upon initial isolation, >90% of all OV stage hPSC populations expressed the definitive neural retinal progenitor marker Vsx2 (Chx10). Comparison of retinal and forebrain progenitor populations revealed key differences in the expression of numerous transcription factors, including Rx, Six6, Dlx1 and Nkx2.1. In vitro maturation of OV populations produced all major classes of retinal cell types in a manner reminiscent of normal development. Furthermore, treatment of hPSC-derived, multipotent early progenitors with factors known to influence retinal development affected cell fate choice in a predictable fashion.

Conclusions: : Results from this study demonstrate that highly enriched populations of OV stage, multipotent retinal progenitors can be isolated from hPSCs. This capability will facilitate future studies of mechanisms of human retinogenesis and disease as well as efforts to develop hPSC-based therapies.

Keywords: retinal development • differentiation • gene/expression 
© 2010, The Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Permission to republish any abstract or part of an abstract in any form must be obtained in writing from the ARVO Office prior to publication.
Copyright © 2024 Association for Research in Vision and Ophthalmology.
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