April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Culture of Polarized Embryonic Stem Cell-Derived RPE on Synthetic Substrates
Danhong Zhu; Sherry T. Hikita; Xuemei Deng; Bo Lu; Lincoln V. Johnson; Dennis O. Clegg; Yuchong Tai; Ashish Ahuja; Mark Humayun; David R. Hinton
Author Affiliations & Notes
  • Danhong Zhu
    Pathology,
    Keck School of Medicine USC, Los Angeles, California
    Doheny Eye Institute, University of Southern California, Los Angeles, California
  • Sherry T. Hikita
    Molecular, Cellular, and Developmental Biology, University of California, Santa Barbara, Santa Barbara, California
  • Xuemei Deng
    Pathology,
    Keck School of Medicine USC, Los Angeles, California
    Doheny Eye Institute, University of Southern California, Los Angeles, California
  • Bo Lu
    Electrical Engineering, California Institute of Technology, Los Angeles, California
  • Lincoln V. Johnson
    Molecular, Cellular, and Developmental Biology, University of California, Santa Barbara, Santa Barbara, California
  • Dennis O. Clegg
    Bioscience II, Univ of California-Santa Barbara, Santa Barbara, California
  • Yuchong Tai
    Electrical Engineering, California Institute of Technology, Los Angeles, California
  • Ashish Ahuja
    Doheny Eye Institute, University of Southern California, Los Angeles, California
  • Mark Humayun
    Ophthalmology,
    Keck School of Medicine USC, Los Angeles, California
    Doheny Eye Institute, University of Southern California, Los Angeles, California
  • David R. Hinton
    Pathology,
    Keck School of Medicine USC, Los Angeles, California
    Doheny Eye Institute, University of Southern California, Los Angeles, California
  • Footnotes
    Commercial Relationships  Danhong Zhu, None; Sherry T. Hikita, None; Xuemei Deng, None; Bo Lu, None; Lincoln V. Johnson, None; Dennis O. Clegg, None; Yuchong Tai, None; Ashish Ahuja, None; Mark Humayun, None; David R. Hinton, None
  • Footnotes
    Support  Grant from CIRM Disease Team Research Award and NIH grants: EY 03040.
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 3191. doi:
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      Danhong Zhu, Sherry T. Hikita, Xuemei Deng, Bo Lu, Lincoln V. Johnson, Dennis O. Clegg, Yuchong Tai, Ashish Ahuja, Mark Humayun, David R. Hinton; Culture of Polarized Embryonic Stem Cell-Derived RPE on Synthetic Substrates. Invest. Ophthalmol. Vis. Sci. 2011;52(14):3191.

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Abstract

Purpose: : RPE cell loss in the macular region is a key pathologic alteration in atrophic age-related macular degeneration (AMD). The restoration of lost RPE through transplantation of a polarized RPE monolayer is a promising new treatment for AMD. RPE cells derived from human embryonic stem cells (hESC-RPE) are a potentially unlimited resource for this therapy. This study tested two synthetic substrates, parylene (non-degradable) and poly(lactic-co-glycolic acid) (PLGA) (bio-degradable) as supports for the growth of polarized hESC-RPE monolayers for potential use in sub-retinal transplantation.

Methods: : 1). hESC-RPE were derived from H9 human embryonic stem cells.2). hESC-RPE were seeded on CellStart-coated (Invitrogen) substrate membranes at 105 cells/cm2 and cultured in XVIVO-10medium (Lonza) for 4 weeks.3). Scanning and transmission electron microscopy were used for ultrastructural analysis of cells and substrates.4).Whole human genome microarray analysis (Agilent) was performed on a parallel culture of hESC-RPE cells, where the cells were seeded on CellStart-coated tissue culture plates and grown in XVIVO-10 medium. RNA was collected at 4 weeks post-seeding.

Results: : hESC-RPE attached, proliferated and differentiated into polarized monolayers on both substrates. They exhibited hexagonal shape, melaninization, and apical microvilli. Microarray results showed that hESC-RPE cells grown in XVIVO10 media expressed signature RPE genes and had higher amounts of PEDF, bestrophin and RDH5, compared with controls.

Conclusions: : Both synthetic substrates supported hESC-RPE cell growth and polarization. They are candidate platforms for differentiation and surgical manipulation of hESC-RPE monolayers for sub-retinal transplantation.

Keywords: age-related macular degeneration • transplantation • retinal pigment epithelium 
© 2011, 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 © 2025 Association for Research in Vision and Ophthalmology.
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