May 2008
Volume 49, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2008
Transplanted Fetal Human Retinal Pigment Epithelium (fRPE) and Human Embryonic Stem Cell-Derived RPE (hES-RPE) Survival on Peripheral Bruch’s Membrane
Author Affiliations & Notes
  • A. C. Johnson
    Ophthalmology, UMDNJ-NJMS, Newark, New Jersey
  • N. Cheewatrakoolpong
    Ophthalmology, UMDNJ-NJMS, Newark, New Jersey
  • C. Fernandes
    Ophthalmology, UMDNJ-NJMS, Newark, New Jersey
  • C. Malcuit
    Advanced Cell Technology Inc., Worcester, Massachusetts
  • L. Vilner
    Advanced Cell Technology Inc., Worcester, Massachusetts
  • L. Lemieux
    Advanced Cell Technology Inc., Worcester, Massachusetts
  • I. Klimanskaya
    Ophthalmology, UMDNJ-NJMS, Newark, New Jersey
  • I. K. Sugino
    Ophthalmology, UMDNJ-NJMS, Newark, New Jersey
  • M. A. Zarbin
    Ophthalmology, UMDNJ-NJMS, Newark, New Jersey
  • Footnotes
    Commercial Relationships  A.C. Johnson, Advanced Cell Technology Inc., F; N. Cheewatrakoolpong, Advanced Cell Technology Inc., F; C. Fernandes, Advanced Cell Technology Inc., F; C. Malcuit, Advanced Cell Technology Inc., E; L. Vilner, Advanced Cell Technology Inc., E; L. Lemieux, Advanced Cell Technology Inc., E; I. Klimanskaya, Advanced Cell Technology Inc., E; I.K. Sugino, Advanced Cell Technology Inc., F; M.A. Zarbin, Advanced Cell Technology Inc., C.
  • Footnotes
    Support  Foundation Fighting Blindness, Research to Prevent Blindness, The Eye Institute of New Jersey, The Janice Mitchell Vassar & Ashby John Mitchell Fellowship, & New Jersey Lions Eye Research Foundation
Investigative Ophthalmology & Visual Science May 2008, Vol.49, 3562. doi:
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      A. C. Johnson, N. Cheewatrakoolpong, C. Fernandes, C. Malcuit, L. Vilner, L. Lemieux, I. Klimanskaya, I. K. Sugino, M. A. Zarbin; Transplanted Fetal Human Retinal Pigment Epithelium (fRPE) and Human Embryonic Stem Cell-Derived RPE (hES-RPE) Survival on Peripheral Bruch’s Membrane. Invest. Ophthalmol. Vis. Sci. 2008;49(13):3562.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : To compare the behavior of fRPE with hES-RPE on peripheral aged human Bruch’s membrane (BM).

Methods: : Explants (N=6 normal (no submacular drusen), N=3 submacular or perimacular drusen; mean age 75.7±9.89 yrs.) were debrided to expose the inner collagenous layer of BM. Fetal RPE or hES-RPE of 3 different degrees of pigmentation (reflecting differing culture conditions) were seeded (~3164 cells/mm2) onto 6mm BM buttons and cultured for 1 or 21 days. Scanning electron microscopy and light microscopy based nuclear density counts were performed on explants. Statistical differences were determined by a Tukey-Kramer Post hoc test after one-way analysis of variance (ANOVA).

Results: : At day-1 after seeding, fRPE showed almost complete coverage of BM while hES-RPE showed few attached cells, regardless of the degree of differentiation. Average nuclear density of fRPE was significantly greater (p<.005) than hES-RPE at day-1. Within the same donor pair, fRPE average nuclear density was significantly greater (p<.005) at day-1 than day-21 in 7/8 explant pairs. hES-RPE preparations had a significantly greater (p<.005) average nuclear density at day-21 compared to day-1 in 9/14 explant pairs. There was no significant difference between 21-day hES-RPE and 21-day fRPE average nuclear density. At day-21, there was poor survival of cells on donor explants from eyes with drusen regardless of cell type or preparation.

Conclusions: : Despite the inability of hES-RPE to attach as rapidly to BM as fRPE, hES-RPE appear to survive on BM to the same degree as fRPE for 21 days. The degree of hES-RPE pigmentation did not appear to have an affect on initial attachment or long term survival.

Keywords: age-related macular degeneration • Bruch's membrane • retinal pigment epithelium 
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