Comparison of FRPE and @entity1 embryonic stem cell-derived RPE behavior on aged XXXX Bruch's membrane.

@entity1 @entity3060 @entity7 @entity18180 @entity21107 @entity13785 @entity2252
PURPOSE: To compare RPE derived from @entity1 embryonic stem cells (hES-RPE) and fetal RPE (fRPE) behavior on @entity1 Bruch's membrane (BM) from aged and @entity13785 donors. METHODS: hES-RPE of 3 degrees of pigmentation and fRPE were cultured on BM explants. Explants were assessed by light, confocal, and scanning electron microscopy. Integrin mRNA levels were determined by real-time polymerase chain reaction studies. Secreted proteins in media were analyzed by multiplex protein analysis after 48-hour exposure at culture day 21. RESULTS: hES-RPE showed impaired initial attachment compared to fRPE; pigmented hES-RPE showed nuclear densities similar to fRPE at day 21. At days 3 and 7, hES-RPE resurfaced BM to a limited degree, showed little proliferation (Ki-67), and partial retention of RPE markers ( @entity3060 , cytokeratin, and @entity21107 ). TUNEL-positive nuclei were abundant at day 3. fRPE exhibited substantial BM resurfacing at day 3 with decreased resurfacing at later times. Most fRPE retained RPE markers. Ki-67-positive nuclei decreased with time in culture. TUNEL staining was variable. Increased integrin mRNA expression did not appear to affect cell survival at day 21. hES-RPE and fRPE protein secretion was similar on equatorial BM except for higher levels of nerve growth factor and @entity18180 ( @entity18180 ) by hES-RPE. On submacular BM, fRPE secreted more @entity7 ( @entity7 ), @entity2252 , and platelet-derived growth factor; hES-RPE secreted more @entity18180 . CONCLUSIONS: Although pigmented hES-RPE and fRPE resurfaced aged and @entity13785 BM to a similar, limited degree at day 21, cell behavior at earlier times was markedly dissimilar. Differences in protein secretion may indicate that hES-RPE may not function identically to native RPE after seeding on aged or @entity13785 BM.
