title: Xeno-Free and Defined Human Embryonic Stem Cell-Derived Retinal Pigment Epithelial Cells Functionally Integrate in a Large-Eyed Preclinical Model.
abstract: Human embryonic stem cell (hESC)-derived retinal pigment epithelial (RPE) cells could replace lost tissue in geographic atrophy (GA) but efficacy has yet to be demonstrated in a large-eyed model. Also, production of hESC-RPE has not yet been achieved in a xeno-free and defined manner, which is critical for clinical compliance and reduced immunogenicity. Here we describe an effective differentiation methodology using human laminin-521 matrix with xeno-free and defined medium. Differentiated cells exhibited characteristics of native RPE including morphology, pigmentation, marker expression, monolayer integrity, and polarization together with phagocytic activity. Furthermore, we established a large-eyed GA model that allowed in vivo imaging of hESC-RPE and host retina. Cells transplanted in suspension showed long-term integration and formed polarized monolayers exhibiting phagocytic and photoreceptor rescue capacity. We have developed a xeno-free and defined hESC-RPE differentiation method and present evidence of functional integration of clinically compliant hESC-RPE in a large-eyed disease model.
authors.affiliation: Department of Clinical Sciences, Intervention and Technology, Karolinska Institutet, 14186 Stockholm, Sweden.
authors.forename: Alvaro
authors.initials: A
authors.lastname: Plaza Reyes
authors.affiliation: Department of Clinical Sciences, Intervention and Technology, Karolinska Institutet, 14186 Stockholm, Sweden; Department of Clinical Neuroscience, Section for Ophthalmology and Vision, St. Erik Eye Hospital, Karolinska Institutet, 11282 Stockholm, Sweden.
authors.forename: Sandra
authors.initials: S
authors.lastname: Petrus-Reurer
authors.affiliation: Department of Clinical Sciences, Intervention and Technology, Karolinska Institutet, 14186 Stockholm, Sweden.
authors.forename: Liselotte
authors.initials: L
authors.lastname: Antonsson
authors.affiliation: Department of Clinical Sciences, Intervention and Technology, Karolinska Institutet, 14186 Stockholm, Sweden.
authors.forename: Sonya
authors.initials: S
authors.lastname: Stenfelt
authors.affiliation: Department of Clinical Neuroscience, Section for Ophthalmology and Vision, St. Erik Eye Hospital, Karolinska Institutet, 11282 Stockholm, Sweden.
authors.forename: Hammurabi
authors.initials: H
authors.lastname: Bartuma
authors.affiliation: Department of Clinical Sciences, Intervention and Technology, Karolinska Institutet, 14186 Stockholm, Sweden.
authors.forename: Sarita
authors.initials: S
authors.lastname: Panula
authors.affiliation: Department of Clinical Sciences, Intervention and Technology, Karolinska Institutet, 14186 Stockholm, Sweden.
authors.forename: Theresa
authors.initials: T
authors.lastname: Mader
authors.affiliation: Department of Medicine, Center for Hematology and Regenerative Medicine, Karolinska Institutet, 14157 Stockholm, Sweden.
authors.forename: Iyadh
authors.initials: I
authors.lastname: Douagi
authors.affiliation: Department of Clinical Neuroscience, Section for Ophthalmology and Vision, St. Erik Eye Hospital, Karolinska Institutet, 11282 Stockholm, Sweden.
authors.forename: Helder
authors.initials: H
authors.lastname: André
authors.affiliation: Department of Clinical Sciences, Intervention and Technology, Karolinska Institutet, 14186 Stockholm, Sweden; Cell Therapy Department, Nova Southeastern University, Fort Lauderdale, FL 33314, USA.
authors.forename: Outi
authors.initials: O
authors.lastname: Hovatta
authors.affiliation: Department of Clinical Sciences, Intervention and Technology, Karolinska Institutet, 14186 Stockholm, Sweden. Electronic address: fredrik.lanner@ki.se.
authors.forename: Fredrik
authors.initials: F
authors.lastname: Lanner
authors.affiliation: Department of Clinical Neuroscience, Section for Ophthalmology and Vision, St. Erik Eye Hospital, Karolinska Institutet, 11282 Stockholm, Sweden.
authors.forename: Anders
authors.initials: A
authors.lastname: Kvanta
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology*; D002460:Cell Line; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D004195:Disease Models, Animal; D057092:Geographic Atrophy / Q000503:physiopathology* / Q000628:therapy; D000066449:Human Embryonic Stem Cells / Q000166:cytology / Q000502:physiology* / Q000637:transplantation; D006801:Humans; D007797:Laminin / Q000378:metabolism; D018613:Microscopy, Confocal; D011817:Rabbits; D055213:Retinal Pigment Epithelium / Q000166:cytology / Q000502:physiology*; D033581:Stem Cell Transplantation / Q000379:methods; D059008:Time-Lapse Imaging; D014183:Transplantation, Heterologous; D015262:Xenobiotics / Q000737:chemistry / Q000494:pharmacology
chemical_list: D003470:Culture Media; D007797:Laminin; D015262:Xenobiotics; C474559:laminin 11 protein, human
keywords: cellular therapy; differentiation; human embryonic stem cells; laminin; optical coherence tomography; rabbit; retinal pigment epithelium; transplantation; xeno-free