A method to enhance cell survival on Bruch's membrane in eyes affected by age and age-related macular degeneration.
OBJECTIVE
To determine whether conditioned medium (CM) derived from bovine corneal endothelial cells (BCECs) can support transplanted cells on aged and age-related macular degeneration (AMD) Bruch's membrane (BM).


METHODS
Retinal pigment epithelium (RPE) cells derived from human embryonic stem cells (hES-RPE) and cultured fetal and aged adult RPE were seeded onto the inner collagenous layer of submacular BM-choroid-sclera explants generated from aged and AMD human donor eyes. Paired explants were cultured in BCEC-CM or CM vehicle. To assess cell behavior after attachment to BM was established, explants were harvested after 21 days in culture. To assess whether sustained exposure to BCEC-CM was necessary for improved cell survival on BM, short exposure to BCEC-CM (3, 7, 14 days) was compared with 21-day exposure. Explants were harvested and evaluated by scanning electron and light microscopy. Extracellular matrix (ECM) deposition after exposure to BCEC-CM was evaluated following RPE cell removal after day 21 on tissue culture dishes or on BM.


RESULTS
BCEC-CM significantly enhanced hES-RPE, fetal RPE, and aged adult RPE survival on BM, regardless of submacular pathology. Although shorter BCEC-CM exposure times showed significant improvement in cell survival compared with culture in CM vehicle, longer BCEC-CM exposure times were more effective. BCEC-CM increased RPE ECM deposition on tissue culture plastic and on BM.


CONCLUSIONS
The results of this study indicate that RPE survival is possible on AMD BM and offer a method that could be developed for enhancing transplanted cell survival on AMD BM. Increased ECM deposition may account for improved cell survival after culture in BCEC-CM.
Institute of Ophthalmology and Visual Science, New Jersey Medical School, University of Medicine and Dentistry of New Jersey, Newark, New Jersey, USA.
Ilene K
IK
Sugino
Aprille
A
Rapista
Qian
Q
Sun
Jianqiu
J
Wang
Celia F
CF
Nunes
Noounanong
N
Cheewatrakoolpong
Marco A
MA
Zarbin
D000368:Aged; D000369:Aged, 80 and over; D000375:Aging / Q000502:physiology*; D000818:Animals; D016570:Bruch Membrane / Q000502:physiology*; D002417:Cattle; D002448:Cell Adhesion / Q000502:physiology; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D024141:Collagen Type IV / Q000378:metabolism; D017077:Culture Media, Conditioned / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology / Q000378:metabolism; D004728:Endothelium, Corneal / Q000166:cytology; D005260:Female; D005333:Fetus / Q000166:cytology; D005353:Fibronectins / Q000378:metabolism; D019084:Fluorescent Antibody Technique, Indirect; D005865:Gestational Age; D006801:Humans; D007797:Laminin / Q000378:metabolism; D008268:Macular Degeneration / Q000378:metabolism*; D008297:Male; D008855:Microscopy, Electron, Scanning; D008875:Middle Aged; D009924:Organ Culture Techniques; D055213:Retinal Pigment Epithelium / Q000166:cytology* / Q000378:metabolism
D024141:Collagen Type IV; D017077:Culture Media, Conditioned; D005353:Fibronectins; D007797:Laminin