J Wassélius1, F Ghosh. 1. Department of Ophthalmology, Lund University Hospital, Sweden.
Abstract
PURPOSE: To study the survival of adult retinal grafts prepared in a physiologically optimized way. METHODS: Twenty-three rabbits received an adult full-thickness rabbit retinal transplant positioned under the host retina, using a vitrectomy technique. The transplants were prepared using a procedure based on a previously described in vitro model used for physiological experiments on the adult retina. Five rabbits received a fragmented graft. All grafts were prelabeled with 4',6-diaminidin-2-phenylindoldihydrochloride (DAPI) to allow identification. The eyes were examined by light and fluorescence microscopy 6 to 174 days after surgery. To assess the amount of cell death in the graft before actual transplantation, in vitro experiments were performed. The extent of cell death in retinas prepared by the optimized protocol was examined and compared with a simpler preparation previously used successfully for embryonic grafts. The amount of cell death in the in vitro experiments was evaluated using a fluorescent green nucleic acid stain that penetrates dying cells. RESULTS: In 21 of the 23 animals that received full-thickness grafts prepared in an optimized way, the transplant survived. Sixteen grafts, including all four with a 174-day survival time, displayed normal morphology, with all retinal layers preserved. The fragmented grafts survived poorly. The in vitro experiments showed minimal cell death in retinas prepared according to the optimized protocol, whereas control retinas displayed extensive cell death after 5 hours. CONCLUSIONS: The results showed that it is possible to transplant adult retina in the rabbit and that the grafts survive well if they are prepared under physiologically optimized conditions and the integrity of the grafted tissue is kept intact.
PURPOSE: To study the survival of adult retinal grafts prepared in a physiologically optimized way. METHODS: Twenty-three rabbits received an adult full-thickness rabbit retinal transplant positioned under the host retina, using a vitrectomy technique. The transplants were prepared using a procedure based on a previously described in vitro model used for physiological experiments on the adult retina. Five rabbits received a fragmented graft. All grafts were prelabeled with 4',6-diaminidin-2-phenylindoldihydrochloride (DAPI) to allow identification. The eyes were examined by light and fluorescence microscopy 6 to 174 days after surgery. To assess the amount of cell death in the graft before actual transplantation, in vitro experiments were performed. The extent of cell death in retinas prepared by the optimized protocol was examined and compared with a simpler preparation previously used successfully for embryonic grafts. The amount of cell death in the in vitro experiments was evaluated using a fluorescent green nucleic acid stain that penetrates dying cells. RESULTS: In 21 of the 23 animals that received full-thickness grafts prepared in an optimized way, the transplant survived. Sixteen grafts, including all four with a 174-day survival time, displayed normal morphology, with all retinal layers preserved. The fragmented grafts survived poorly. The in vitro experiments showed minimal cell death in retinas prepared according to the optimized protocol, whereas control retinas displayed extensive cell death after 5 hours. CONCLUSIONS: The results showed that it is possible to transplant adult retina in the rabbit and that the grafts survive well if they are prepared under physiologically optimized conditions and the integrity of the grafted tissue is kept intact.
Authors: Fredrik Ghosh; Karl Engelsberg; Robert V English; Robert M Petters Journal: Graefes Arch Clin Exp Ophthalmol Date: 2006-10-27 Impact factor: 3.535
Authors: Tai-Chi Lin; Magdalene J Seiler; Danhong Zhu; Paulo Falabella; David R Hinton; Dennis O Clegg; Mark S Humayun; Biju B Thomas Journal: Stem Cells Int Date: 2017-08-27 Impact factor: 5.443