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