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L. Geng, L.V. Del Priore, T.H. Tezel, H.J. Kaplan; RPE Sheet Transplantation in Porcine Eyes: Evidence for Basement Membrane Synthesis and Long-Term Cell Survival . Invest. Ophthalmol. Vis. Sci. 2003;44(13):506.
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Purpose: To determine the survival and morphologic appearance after transplantation of organized primary porcine RPE sheets into the subretinal space of the pig eye. Methods: Sheets of primary female RPE cells were harvested from freshly enucleated porcine eyes by removing the sclera and incubating the eyecup with 25 U/ml of Dispase for 30 minutes. RPE viability was determined with an esterase stain. RPE sheets were transplanted into the subretinal space of male pigs using pars plana vitrectomy techniques. 38 animals underwent surgery and the transplant bed was examined by light and transmission electron microscopy up to 3 months later. Transplanted RPE cells were identified by expression of Barr body. Results: The current study demonstrates that transplanted cells can be identified using sex chromatin. Four days after surgery there was some folding of the transplant with heavily pigmented RPE cells that were predominantly Barr body positive. One month after surgery the transplant appeared as a monolayer in some regions and multilayer in other regions. There was evidence of basement membrane synthesis by the transplanted sheets. Barr body positive cells could still be identified 3 months after surgery, although there were many Barr body negative pigment-laden cells that may represent host RPE or macrophages in the subretinal space. The major change seen in the retina after surgery was shortening of the photoreceptor outer segments during the first month after surgery, with recovery to normal length by 3 months. The choroidal vessels and choriocapillaris remained patent in the transplant bed up to 3 months. Some inflammatory cells were visible within the vessels of the choriocapillaris 2-4 weeks after surgery. Conclusions: Transplanted RPE sheets survive in the subretinal space up to three months after surgery. Adjacent choriocapillaris remains patent under the transplant. Some of the heavily pigmented cells within the transplant bed are Barr body negative macrophages at 3 months after surgery. RPE aligned basally along Bruch's membrane maintain a better morphology, in contrast to folded inner layers of transplanted RPE sheet, which demonstrate degeneration within one month after surgery.
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