title: Retinal pigmented epithelial cells obtained from human induced pluripotent stem cells possess functional visual cycle enzymes in vitro and in vivo.
abstract: Differentiated retinal pigmented epithelial (RPE) cells have been obtained from human induced pluripotent stem (hiPS) cells. However, the visual (retinoid) cycle in hiPS-RPE cells has not been adequately examined. Here we determined the expression of functional visual cycle enzymes in hiPS-RPE cells compared with that of isolated wild-type mouse primary RPE (mpRPE) cells in vitro and in vivo. hiPS-RPE cells appeared morphologically similar to mpRPE cells. Notably, expression of certain visual cycle proteins was maintained during cell culture of hiPS-RPE cells, whereas expression of these same molecules rapidly decreased in mpRPE cells. Production of the visual chromophore, 11-cis-retinal, and retinosome formation also were documented in hiPS-RPE cells in vitro. When mpRPE cells with luciferase activity were transplanted into the subretinal space of mice, bioluminance intensity was preserved for >3 months. Additionally, transplantation of mpRPE into blind Lrat(-/-) and Rpe65(-/-) mice resulted in the recovery of visual function, including increased electrographic signaling and endogenous 11-cis-retinal production. Finally, when hiPS-RPE cells were transplanted into the subretinal space of Lrat(-/-) and Rpe65(-/-) mice, their vision improved as well. Moreover, histological analyses of these eyes displayed replacement of dysfunctional RPE cells by hiPS-RPE cells. Together, our results show that hiPS-RPE cells can exhibit a functional visual cycle in vitro and in vivo. These cells could provide potential treatment options for certain blinding retinal degenerative diseases.
authors.affiliation: From the Departments of Ophthalmology and Visual Sciences.
authors.forename: Tadao
authors.initials: T
authors.lastname: Maeda
authors.forename: Mee Jee
authors.initials: MJ
authors.lastname: Lee
authors.forename: Grazyna
authors.initials: G
authors.lastname: Palczewska
authors.forename: Stefania
authors.initials: S
authors.lastname: Marsili
authors.forename: Paul J
authors.initials: PJ
authors.lastname: Tesar
authors.forename: Krzysztof
authors.initials: K
authors.lastname: Palczewski
authors.forename: Masayo
authors.initials: M
authors.lastname: Takahashi
authors.forename: Akiko
authors.initials: A
authors.lastname: Maeda
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D002478:Cells, Cultured; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000201:enzymology / Q000637:transplantation*; D051379:Mice; D012162:Retinal Degeneration / Q000235:genetics* / Q000473:pathology / Q000628:therapy*; D055213:Retinal Pigment Epithelium / Q000201:enzymology / Q000637:transplantation*; D012172:Retinaldehyde / Q000096:biosynthesis / Q000235:genetics; D014785:Vision, Ocular / Q000235:genetics / Q000502:physiology; D019745:cis-trans-Isomerases / Q000172:deficiency / Q000235:genetics*; D000099178:Retinoid Isomerohydrolase
chemical_list: D012172:Retinaldehyde; D019745:cis-trans-Isomerases; D000099178:Retinoid Isomerohydrolase
keywords: Induced Pluripotent Stem (iPS) Cell; LRAT; RPE; RPE65; Retinal Degeneration; Retinal Metabolism; Retinoid; Transplantation; Visual Cycle