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T. Abe, M. Hojo, Y. Saigo, E. Sugano, T. Kano, Y. Yoshioka, A. Yamashita, M. Tamai, M. Yamato, M. Okano; Pigment Epithelial Cell Transplantation Recovered From Thermally Responsive Polymer-Grafted Surface . Invest. Ophthalmol. Vis. Sci. 2003;44(13):494.
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Purpose: To prepare the retinal pigment epithelial (RPE) or iris pigment epithlial (IPE) cell from the thermally responsive polymer (poly-(N-isopropylacrylamide))-grafted culture plates and to examine its effect for transplantation. Methods: RPE and IPE were prepared from Long Evans rats and monkeys, respectively. These cells were cultured on thermally responsive polymer-grafted culture plates or on normal culture plates. These cells were recovered from the culture plates either by placing the cells from 37oC to 20oC or by trypsin treatment. Western blotting analysis and enzyme linked immunosorbent assay (ELISA) were performed against chondroitin sulfate or collagen type IV, respectively. These cells were also transplanted into subretinal regions of three monkeys. Histological and electron microscopical examinations were performed. Results: Each cell was recovered as cell sheet from thermally responsive polymer-grafted culture plates by changing the temperature from 37oC to 20oC or as cell suspension using trypsin. When we performed western blotting analysis or ELISA, we found that chondroitin sulfate or collagen type IV showed higher concentration in the cells recovered by thermally responsive culture plates than those recovered by trypsin treatment. Histological and electron microscopical examination of the transplanted region of the monkeys revealed that the cells transplanted by sheet embraced the photoreceptor outer segments more than that of the cells recovered by trypsin treatment. Electron microscopical examination also showed tight junction-like structures between the transplanted cells. Conclusions: Pigment epithelial cells recovered by thermally responsive polymer-grafted culture plates had more extracellular matrix proteins than that of the cells recovered by trypsin treatment. Transplantation of these cells may have more functional properties than that of the cells recovered by trypsin.
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