title: Generation of retinal pigment epithelial cells from small molecules and OCT4 reprogrammed human induced pluripotent stem cells.
abstract: Autologous retinal pigment epithelium (RPE) grafts derived from induced pluripotent stem cells (iPSCs) may be used to cure blinding diseases in which RPE dysfunction results in photoreceptor degeneration. Four-, two-, and one-factor-derived iPSCs (4F-, 2F-, and 1F-iPSCs, respectively) were differentiated into fully functional cuboidal pigmented cells in polarized monolayers that express RPE-specific markers. 1F-iPSCs-RPE (1F-iPS-RPE) strongly resembles primary human fetal RPE (hfRPE) based on proteomic and untargeted metabolomic analyses, and using novel in vivo imaging technology coupled with electroretinography, we demonstrated that 1F-iPS-RPE mediate anatomical and functional rescue of photoreceptors after transplantation in an animal model of RPE-mediated retinal degeneration. 1F-iPS0RPE cells were injected subretinally as a suspension and formed a monolayer dispersed between host RPE cells. Furthermore, 1F-iPS-RPE do not simply provide trophic support to rescue photoreceptors as previously speculated but actually phagocytose photoreceptor outer segments in vivo and maintain visual cycling. Thus, 1f-iPS-RPE grafts may be superior to conventional iPS-RPE for clinical use because 1F-IPS-RPE closely resemble hfRPE, mediate anatomical and functional photoreceptor rescue in vivo, and are generated using a reduced number of potentially oncogenic reprogramming factors.
authors.affiliation: Department of Cell Biology, The Scripps Research Institute, La Jolla, California 92037, USA.
authors.forename: Tim U
authors.initials: TU
authors.lastname: Krohne
authors.forename: Peter D
authors.initials: PD
authors.lastname: Westenskow
authors.forename: Toshihide
authors.initials: T
authors.lastname: Kurihara
authors.forename: David F
authors.initials: DF
authors.lastname: Friedlander
authors.forename: Mandy
authors.initials: M
authors.lastname: Lehmann
authors.forename: Alison L
authors.initials: AL
authors.lastname: Dorsey
authors.forename: Wenlin
authors.initials: W
authors.lastname: Li
authors.forename: Saiyong
authors.initials: S
authors.lastname: Zhu
authors.forename: Andrew
authors.initials: A
authors.lastname: Schultz
authors.forename: Junhua
authors.initials: J
authors.lastname: Wang
authors.forename: Gary
authors.initials: G
authors.lastname: Siuzdak
authors.forename: Sheng
authors.initials: S
authors.lastname: Ding
authors.forename: Martin
authors.initials: M
authors.lastname: Friedlander
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D017077:Culture Media, Conditioned; D004596:Electroretinography; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D015603:Keratinocytes / Q000166:cytology / Q000378:metabolism; D055432:Metabolomics; D023421:Models, Animal; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3 / Q000235:genetics / Q000378:metabolism*; D010587:Phagocytosis; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D051381:Rats; D012162:Retinal Degeneration / Q000378:metabolism / Q000628:therapy; D055214:Retinal Photoreceptor Cell Outer Segment / Q000378:metabolism / Q000637:transplantation; D055213:Retinal Pigment Epithelium / Q000166:cytology* / Q000378:metabolism; D013552:Swine / Q000378:metabolism
chemical_list: D015415:Biomarkers; D017077:Culture Media, Conditioned; D018398:Homeodomain Proteins; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D050814:Octamer Transcription Factor-3; C494173:POU5F1 protein, human; D012333:RNA, Messenger