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K. Ohno–Matsui, K. Mori, S. Ichinose, T. Sato, A. Kojima, N. Shimada, I. Morita, M. Mochizuki; Photoreceptor Protection by Iris Pigment Epithelial Transplantation Cultured on Amniotic Membrane . Invest. Ophthalmol. Vis. Sci. 2006;47(13):1400.
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© ARVO (1962-2015); The Authors (2016-present)
To examine the characteristics of iris pigment epithelial (IPE) cells cultured on amniotic membrane (AM) and to determine whether subretinal transplantation of this IPE cell sheet can protect photoreceptor cell loss in dystrophic Royal College of Surgeons (RCS) rats.
Human and rat IPE cells were cultured on the basement membrane side of dispase treated AM. After 2 weeks of seeding, the expression of several genes participate in the function of differentiated retinal pigment epithelial (RPE) cells was evaluated using real time PCR and Western blotting. Ultrastructural changes were evaluated using transmission electron microscopy. The IPE cell sheet cultured on AM was transplanted into the subretinal space of 4–week–old RCS rats, and the eyes were analyzed histologically at 12 weeks after grafting. The effect of transplantation of IPE cell sheet on the visual function was estimated by optokinetic reflex.
IPE cells cultured on AM exhibited ultrastructural epithelial features such as apical microvilli and intercellular junctions. The gene expression of differentiation markers of RPE cells in IPE cells on AM, compared with those of IPE cells cultured on uncoated plastic dishes, was up–regulated; PEDF was markedly and RPE65, bestrophin, VEGF, and BDNF were slightly. In contrast, CRALBP expression was not changed. A statistically significant photoreceptor protection was observed in RCS rats which received subretinal transplantation of IPE cell sheet on AM 12 weeks after operation compared to nontreated dystrophic rats. When screened for head–tracking to moving stripes, IPE cell sheet–grafted animals performed better than sham–treated or control dystrophic animals.
Subretinal transplantation of IPE cells sheet cultivated on AM limit functional deterioration and prolong photoreceptor survival in a rat model of retinal degeneration. This suggests that AM might be a useful matrix substrate for IPE cell transplantation, however, more modification might be necessary to fully reproduce differentiated features of RPE cells.
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