PURPOSE: To study topographical differences in porcine retinal pigment epithelial (RPE) cell proliferation (1) in vivo, after experimental central surgical subretinal injury, and (2) in vitro. METHODS: Domestic pigs underwent either experimental RPE debridement (n = 5), subretinal amniotic membrane transplantation (n= 4), or both (n= 1) in the left eye. RPE cell proliferation was assayed by injection of the thymidine analogue 5-bromodeoxyuridine (5-BrdU) at postoperative day 0 and 1. RPE cells in S-phase were identified by their incorporation of 5-BrdU, as detected by immunohistochemistry. The in vitro proliferation of primary RPE isolates from the peripheral and central retina was assayed by a colorimetric assay and by [(3)H]thymidine incorporation. RESULTS: After subretinal surgery, in vivo incorporation of 5-BrdU was seen in peripheral RPE cells in 8 of 10 surgically treated eyes, but never in central RPE cells. This observation was true of both types of experimental surgery performed. In vitro, RPE isolates from the pre-equatorial region consistently yielded higher cell densities than did RPE cell isolates from more central parts of the epithelium. This was apparent through the three first passages of porcine RPE cells in culture. After 1 and 4 days in culture, pre-equatorial RPE cells had incorporated significantly more [(3)H]thymidine than had the more central RPE cells. CONCLUSIONS: Experimental subretinal surgical injury of the RPE below the central retina is followed within 48 hours by a peripheral, but not a central, proliferation of RPE cells. In vitro, peripheral RPE cells have a higher proliferative capacity than do central RPE cells.
PURPOSE: To study topographical differences in porcine retinal pigment epithelial (RPE) cell proliferation (1) in vivo, after experimental central surgical subretinal injury, and (2) in vitro. METHODS:Domestic pigs underwent either experimental RPE debridement (n = 5), subretinal amniotic membrane transplantation (n= 4), or both (n= 1) in the left eye. RPE cell proliferation was assayed by injection of the thymidine analogue 5-bromodeoxyuridine (5-BrdU) at postoperative day 0 and 1. RPE cells in S-phase were identified by their incorporation of 5-BrdU, as detected by immunohistochemistry. The in vitro proliferation of primary RPE isolates from the peripheral and central retina was assayed by a colorimetric assay and by [(3)H]thymidine incorporation. RESULTS: After subretinal surgery, in vivo incorporation of 5-BrdU was seen in peripheral RPE cells in 8 of 10 surgically treated eyes, but never in central RPE cells. This observation was true of both types of experimental surgery performed. In vitro, RPE isolates from the pre-equatorial region consistently yielded higher cell densities than did RPE cell isolates from more central parts of the epithelium. This was apparent through the three first passages of porcine RPE cells in culture. After 1 and 4 days in culture, pre-equatorial RPE cells had incorporated significantly more [(3)H]thymidine than had the more central RPE cells. CONCLUSIONS: Experimental subretinal surgical injury of the RPE below the central retina is followed within 48 hours by a peripheral, but not a central, proliferation of RPE cells. In vitro, peripheral RPE cells have a higher proliferative capacity than do central RPE cells.
Authors: Michael J Koss; Paulo Falabella; Francisco R Stefanini; Marcel Pfister; Biju B Thomas; Amir H Kashani; Rodrigo Brant; Danhong Zhu; Dennis O Clegg; David R Hinton; Mark S Humayun Journal: Graefes Arch Clin Exp Ophthalmol Date: 2016-06-22 Impact factor: 3.117