DOI: 10.1167/tvst.5.1.8 Article Two Bioactive Molecular Weight Fractions of a Conditioned Medium Enhance RPE Cell Survival on Age-Related Macular Degeneration and Aged Bruch’s Membrane Ilene K. Sugino1, Qian Sun1, Carola Springer1, Noounanong Cheewatrakoolpong1, Tong Liu2, Hong Li2, and Marco A. Zarbin1 1 Institute of Ophthalmology and Visual Science, Rutgers, New Jersey Medical School, Newark, NJ, USA 2 Department of Biochemistry and Molecular Biology, Center for Advanced Proteomics Research, Neuroproteomics Core Facility, Rutgers, New Jersey Medical School, Newark, NJ, USA Correspondence: Marco A. Zarbin, Purpose: To characterize molecular weight fractions of bovine corneal endothelial cell Institute of Ophthalmology and conditioned medium (CM) supporting retinal pigment epithelium (RPE) cell survival Visual Science, 90 Bergen Street, on aged and age-related macular degeneration (AMD) Bruch’s membrane. Newark, NJ 07101-1709, USA;
[email protected] Methods: CM was subject to size separation using centrifugal filters. Retentate and filtrate fractions were tested for bioactivity by analyzing RPE survival on submacular Received: 4 September 2015 Bruch’s membrane of aged and AMD donor eyes and behavior on collagen I-coated Accepted: 1 January 2016 tissue culture wells. Protein and peptide composition of active fractions was Published: 22 February 2016 determined by mass spectrometry. Keywords: age-related macular Results: Two bioactive fractions, 3-kDa filtrate and a 10-50–kDa fraction, were degeneration; retinal pigment ep- necessary for RPE survival on aged and AMD Bruch’s membrane. The 3-kDa filtrate, ithelium; cell transplantation; but not the 10-50–kDa fraction, supported RPE growth on collagen 1-coated tissue Bruch’s membrane; cell survival culture plates.