A method to enhance cell survival on Bruch's membrane in eyes affected by age and age-related macular degeneration

• Published in: Investigative ophthalmology & visual science Vol. 52; no. 13; pp. 9598 - 9609
• Main Authors: Sugino, Ilene K, Rapista, Aprille, Sun, Qian, Wang, Jianqiu, Nunes, Celia F, Cheewatrakoolpong, Noounanong, Zarbin, Marco A
• Format: Journal Article
• Language: English
• Published: United States 20-12-2011
• Subjects: Aged; Aged, 80 and over; Aging - physiology; Animals; Bruch Membrane - physiology; Cattle; Cell Adhesion - physiology; Cell Survival - physiology; Cells, Cultured; Collagen Type IV - metabolism; Culture Media, Conditioned - pharmacology; Embryonic Stem Cells - cytology; Embryonic Stem Cells - metabolism; Endothelium, Corneal - cytology; Female; Fetus - cytology; Fibronectins - metabolism; Fluorescent Antibody Technique, Indirect; Gestational Age; Humans; Laminin - metabolism; Macular Degeneration - metabolism; Male; Microscopy, Electron, Scanning; Middle Aged; Organ Culture Techniques; Retinal Pigment Epithelium - cytology; Retinal Pigment Epithelium - metabolism
• ISSN: 1552-5783; 1552-5783
• DOI: 10.1167/iovs.11-8400

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).
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.
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.
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.