PURPOSE: To determine whether resurfacing submacular human Bruch's membrane with a cell-deposited extracellular matrix (ECM) improves retinal pigment epithelial (RPE) survival. METHODS: Bovine corneal endothelial (BCE) cells were seeded onto the inner collagenous layer of submacular Bruch's membrane explants of human donor eyes to allow ECM deposition. Control explants from fellow eyes were cultured in medium only. The deposited ECM was exposed by removing BCE. Fetal RPE cells were then cultured on these explants for 1, 14, or 21 days. The explants were analyzed quantitatively by light microscopy and scanning electron microscopy. Surviving RPE cells from explants cultured for 21 days were harvested to compare bestrophin and RPE65 mRNA expression. Mass spectroscopy was performed on BCE-ECM to examine the protein composition. RESULTS: The BCE-treated explants showed significantly higher RPE nuclear density than did the control explants at all time points. RPE expressed more differentiated features on BCE-treated explants than on untreated explants, but expressed very little mRNA for bestrophin or RPE65. The untreated young (<50 years) and African American submacular Bruch's membrane explants supported significantly higher RPE nuclear densities (NDs) than did the Caucasian explants. These differences were reduced or nonexistent in the BCE-ECM-treated explants. Proteins identified in the BCE-ECM included ECM proteins, ECM-associated proteins, cell membrane proteins, and intracellular proteins. CONCLUSIONS: Increased RPE survival can be achieved on aged submacular human Bruch's membrane by resurfacing the latter with a cell-deposited ECM. Caucasian eyes seem to benefit the most, as cell survival is the worst on submacular Bruch's membrane in these eyes.
PURPOSE: To determine whether resurfacing submacular human Bruch's membrane with a cell-deposited extracellular matrix (ECM) improves retinal pigment epithelial (RPE) survival. METHODS:Bovine corneal endothelial (BCE) cells were seeded onto the inner collagenous layer of submacular Bruch's membrane explants of humandonor eyes to allow ECM deposition. Control explants from fellow eyes were cultured in medium only. The deposited ECM was exposed by removing BCE. Fetal RPE cells were then cultured on these explants for 1, 14, or 21 days. The explants were analyzed quantitatively by light microscopy and scanning electron microscopy. Surviving RPE cells from explants cultured for 21 days were harvested to compare bestrophin and RPE65 mRNA expression. Mass spectroscopy was performed on BCE-ECM to examine the protein composition. RESULTS: The BCE-treated explants showed significantly higher RPE nuclear density than did the control explants at all time points. RPE expressed more differentiated features on BCE-treated explants than on untreated explants, but expressed very little mRNA for bestrophin or RPE65. The untreated young (<50 years) and African American submacular Bruch's membrane explants supported significantly higher RPE nuclear densities (NDs) than did the Caucasian explants. These differences were reduced or nonexistent in the BCE-ECM-treated explants. Proteins identified in the BCE-ECM included ECM proteins, ECM-associated proteins, cell membrane proteins, and intracellular proteins. CONCLUSIONS: Increased RPE survival can be achieved on aged submacular human Bruch's membrane by resurfacing the latter with a cell-deposited ECM. Caucasian eyes seem to benefit the most, as cell survival is the worst on submacular Bruch's membrane in these eyes.
Authors: Yvette P Conley; Johanna Jakobsdottir; Tammy Mah; Daniel E Weeks; Ronald Klein; Lewis Kuller; Robert E Ferrell; Michael B Gorin Journal: Hum Mol Genet Date: 2006-09-25 Impact factor: 6.150
Authors: Antonia M Joussen; Florian M A Heussen; Sandra Joeres; Helene Llacer; Beate Prinz; Klaus Rohrschneider; Kristel J M Maaijwee; Jan van Meurs; Bernd Kirchhof Journal: Am J Ophthalmol Date: 2006-07 Impact factor: 5.258
Authors: Silke Schmidt; Michael A Hauser; William K Scott; Eric A Postel; Anita Agarwal; Paul Gallins; Frank Wong; Yu Sarah Chen; Kylee Spencer; Nathalie Schnetz-Boutaud; Jonathan L Haines; Margaret A Pericak-Vance Journal: Am J Hum Genet Date: 2006-03-20 Impact factor: 11.025
Authors: David M Brown; Peter K Kaiser; Mark Michels; Gisele Soubrane; Jeffrey S Heier; Robert Y Kim; Judy P Sy; Susan Schneider Journal: N Engl J Med Date: 2006-10-05 Impact factor: 91.245
Authors: Philip J Rosenfeld; David M Brown; Jeffrey S Heier; David S Boyer; Peter K Kaiser; Carol Y Chung; Robert Y Kim Journal: N Engl J Med Date: 2006-10-05 Impact factor: 91.245
Authors: Eunkyung An; Xiaoning Lu; Jessica Flippin; Joseph M Devaney; Brian Halligan; Eric P Hoffman; Eric Hoffman; Nataly Strunnikova; Karl Csaky; Yetrib Hathout Journal: J Proteome Res Date: 2006-10 Impact factor: 4.466
Authors: Anni Sorkio; Heidi Hongisto; Kai Kaarniranta; Hannu Uusitalo; Kati Juuti-Uusitalo; Heli Skottman Journal: Tissue Eng Part A Date: 2014-01-20 Impact factor: 3.845
Authors: Ilene K Sugino; Qian Sun; Jianqiu Wang; Celia F Nunes; Noounanong Cheewatrakoolpong; Aprille Rapista; Adam C Johnson; Christopher Malcuit; Irina Klimanskaya; Robert Lanza; Marco A Zarbin Journal: Invest Ophthalmol Vis Sci Date: 2011-07-01 Impact factor: 4.799
Authors: John C Dreixler; Jacqueline N Poston; Irina Balyasnikova; Afzhal R Shaikh; Kelsey Y Tupper; Sineadh Conway; Venkat Boddapati; Marcus M Marcet; Maciej S Lesniak; Steven Roth Journal: Invest Ophthalmol Vis Sci Date: 2014-04-03 Impact factor: 4.799
Authors: Kathryn L McCabe; Noelia J Kunzevitzky; Brian P Chiswell; Xin Xia; Jeffrey L Goldberg; Robert Lanza Journal: PLoS One Date: 2015-12-21 Impact factor: 3.240