J Clin Ophthalmol Eye Disord | Volume 1, Issue 1 | Research Article | Open Access
Dan-Ning Hu1,2, Ronald C Gentile1, Steven A McCormick1, Pei-Yu Yang2, Thomas O Muldoon1, Hung-Yu Lin2, Uri R Shabto1, Jeanne L Rosenthal1 and Richard B Rosen1*
1Department of Ophthalmology, New York Eye and Ear Infirmary, New York, USA
2Department of Ophthalmology, Show Chwan Memorial Hospital, Changhua, Taiwan
*Correspondance to: Richard B Rosen
Fulltext PDFObjectives: There were few studies on the culture of cells from surgical excised pre- or sub-retinal membranes from Proliferative Vitreoretinopathy (PVR) and Age-Related Macular Degeneration (AMD) patients and all of these studies were restricted to primary cultures only. The purpose of this study was to culture and identify various cellular components of surgically excised membranes from these patients and to explore the role of these cells in the pathogenesis of PVR and AMD by the investigation of the in vitro behavior of these cells in and beyond the primary culture.
Methods: Surgically excised membranes were plated in culture dishes by an explants method and cultured with F12 medium containing fetal bovine serum. Immunocytochemical staining was performed to identify cell types. After the confluence of primary culture, cells were subcultured until senescence.
Results: Of the membranes from 41 PVR eyes, 29 cases exhibited cell outgrowth (71%), and 13 cases (32%) grew to confluence. RPE cells, glial cells, fibroblasts and macrophages were the main cell types identified in the outgrowth of these membranes. RPE cells were the predominant and most actively growing cell type and the sole cell type persisted after second subcultures. RPE cells could be cultured up to 12 generations with 30 divisions in vitro. Of the membranes from 16 AMD eyes, 5 (31%) demonstrated cell outgrowth, but none of these cultures became confluent. Conclusions: Cells from PVR membranes exhibited active proliferation and were predominantly RPE cells. Cells from sub-retinal membranes of AMD eyes showed poor viability and growth capacity. These results were consistent with the pathologic and cell biologic changes of RPE cells in the AMD and PVR.
Age-related macular degeneration; Cell culture; Pre-retinal membrane; Proliferative vitreoretinopathy
Hu D-N, Gentile RC, McCormick SA, Yang P-Y, Muldoon TO, Lin H-Y, et al. Role of RPE Cells in Pathogenesis of Proliferative Vitreoretinopathy and Age-Related Macular Degeneration: Cell Culture Study of Surgical Excised Pre- and Sub-Retinal Membranes. J Clin Ophthalmol Eye Disord. 2017; 1(1): 1002.