title: Cell-deposited matrix improves retinal pigment epithelium survival on aged submacular human Bruch's membrane.
abstract: OBJECTIVE
  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.
authors.affiliation: The Institute of Ophthalmology and Visual Science, New Jersey Medical School, University of Medicine and Dentistry of New Jersey, Newark, New Jersey 07101-1709, USA.
authors.forename: Ilene K
authors.initials: IK
authors.lastname: Sugino
authors.forename: Vamsi K
authors.initials: VK
authors.lastname: Gullapalli
authors.forename: Qian
authors.initials: Q
authors.lastname: Sun
authors.forename: Jianqiu
authors.initials: J
authors.lastname: Wang
authors.forename: Celia F
authors.initials: CF
authors.lastname: Nunes
authors.forename: Noounanong
authors.initials: N
authors.lastname: Cheewatrakoolpong
authors.forename: Adam C
authors.initials: AC
authors.lastname: Johnson
authors.forename: Benjamin C
authors.initials: BC
authors.lastname: Degner
authors.forename: Jianyuan
authors.initials: J
authors.lastname: Hua
authors.forename: Tong
authors.initials: T
authors.lastname: Liu
authors.forename: Wei
authors.initials: W
authors.lastname: Chen
authors.forename: Hong
authors.initials: H
authors.lastname: Li
authors.forename: Marco A
authors.initials: MA
authors.lastname: Zarbin
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000375:Aging / Q000502:physiology*; D000818:Animals; D000075002:Bestrophins; D016570:Bruch Membrane / Q000502:physiology* / Q000648:ultrastructure; D002352:Carrier Proteins / Q000235:genetics; D002417:Cattle; D002452:Cell Count; D018929:Cell Culture Techniques; D002470:Cell Survival / Q000502:physiology; D018118:Chloride Channels / Q000235:genetics; D018920:Coculture Techniques; D004728:Endothelium, Corneal / Q000166:cytology / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism*; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005136:Eye Proteins / Q000235:genetics; D005260:Female; D006801:Humans; D008297:Male; D013058:Mass Spectrometry; D008855:Microscopy, Electron, Scanning; D008875:Middle Aged; D009924:Organ Culture Techniques; D012333:RNA, Messenger / Q000235:genetics; D055213:Retinal Pigment Epithelium / Q000166:cytology* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D019745:cis-trans-Isomerases; D000099178:Retinoid Isomerohydrolase
chemical_list: D000075002:Bestrophins; D002352:Carrier Proteins; D018118:Chloride Channels; D016326:Extracellular Matrix Proteins; D005136:Eye Proteins; D012333:RNA, Messenger; D019745:cis-trans-Isomerases; D000099178:Retinoid Isomerohydrolase; C113133:BEST1 protein, human