title: In Pursuit of Authenticity: Induced Pluripotent Stem Cell-Derived Retinal Pigment Epithelium for Clinical Applications.
abstract: : Induced pluripotent stem cells (iPSCs) can be efficiently differentiated into retinal pigment epithelium (RPE), offering the possibility of autologous cell replacement therapy for retinal degeneration stemming from RPE loss. The generation and maintenance of epithelial apical-basolateral polarity is fundamental for iPSC-derived RPE (iPSC-RPE) to recapitulate native RPE structure and function. Presently, no criteria have been established to determine clonal or donor based heterogeneity in the polarization and maturation state of iPSC-RPE. We provide an unbiased structural, molecular, and physiological evaluation of 15 iPSC-RPE that have been derived from distinct tissues from several different donors. We assessed the intact RPE monolayer in terms of an ATP-dependent signaling pathway that drives critical aspects of RPE function, including calcium and electrophysiological responses, as well as steady-state fluid transport. These responses have key in vivo counterparts that together help determine the homeostasis of the distal retina. We characterized the donor and clonal variation and found that iPSC-RPE function was more significantly affected by the genetic differences between different donors than the epigenetic differences associated with different starting tissues. This study provides a reference dataset to authenticate genetically diverse iPSC-RPE derived for clinical applications.
  
  
  CONCLUSIONS
  The retinal pigment epithelium (RPE) is essential for maintaining visual function. RPE derived from human induced pluripotent stem cells (iPSC-RPE) offer a promising cell-based transplantation therapy for slowing or rescuing RPE-induced visual function loss. For effective treatment, iPSC-RPE must recapitulate the physiology of native human RPE. A set of physiologically relevant functional assays are provided that assess the polarized functional activity and maturation state of the intact RPE monolayer. The present data show that donor-to-donor variability exceeds the tissue-to-tissue variability for a given donor and provides, for the first time, criteria necessary to identify iPSC-RPE most suitable for clinical application.
authors.affiliation: Section on Epithelial and Retinal Physiology and Disease, National Eye Institute, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Kiyoharu J
authors.initials: KJ
authors.lastname: Miyagishima
authors.affiliation: Section on Epithelial and Retinal Physiology and Disease, National Eye Institute, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Qin
authors.initials: Q
authors.lastname: Wan
authors.affiliation: Columbia Stem Cell Core Facility, Columbia University Medical Center, New York, New York, USA.
authors.forename: Barbara
authors.initials: B
authors.lastname: Corneo
authors.affiliation: Unit on Ocular and Stem Cell Translational Research, National Eye Institute, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Ruchi
authors.initials: R
authors.lastname: Sharma
authors.affiliation: Section on Epithelial and Retinal Physiology and Disease, National Eye Institute, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Mostafa R
authors.initials: MR
authors.lastname: Lotfi
authors.affiliation: Neural Stem Cell Institute, Rensselaer, New York, USA.
authors.forename: Nathan C
authors.initials: NC
authors.lastname: Boles
authors.affiliation: Section on Epithelial and Retinal Physiology and Disease, National Eye Institute, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Fang
authors.initials: F
authors.lastname: Hua
authors.affiliation: Section on Epithelial and Retinal Physiology and Disease, National Eye Institute, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Arvydas
authors.initials: A
authors.lastname: Maminishkis
authors.affiliation: Section on Epithelial and Retinal Physiology and Disease, National Eye Institute, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Congxiao
authors.initials: C
authors.lastname: Zhang
authors.affiliation: Department of Development and Regenerative Biology, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
authors.forename: Timothy
authors.initials: T
authors.lastname: Blenkinsop
authors.affiliation: Section on Epithelial and Retinal Physiology and Disease, National Eye Institute, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Vladimir
authors.initials: V
authors.lastname: Khristov
authors.affiliation: Unit on Ocular and Stem Cell Translational Research, National Eye Institute, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Balendu S
authors.initials: BS
authors.lastname: Jha
authors.affiliation: Section on Epithelial and Retinal Physiology and Disease, National Eye Institute, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Omar S
authors.initials: OS
authors.lastname: Memon
authors.affiliation: Department of Pharmacology and Systems Therapeutics, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
authors.forename: Sunita
authors.initials: S
authors.lastname: D'Souza
authors.affiliation: Neural Stem Cell Institute, Rensselaer, New York, USA.
authors.forename: Sally
authors.initials: S
authors.lastname: Temple
authors.affiliation: Section on Epithelial and Retinal Physiology and Disease, National Eye Institute, National Institutes of Health, Bethesda, Maryland, USA.
authors.forename: Sheldon S
authors.initials: SS
authors.lastname: Miller
authors.affiliation: Unit on Ocular and Stem Cell Translational Research, National Eye Institute, National Institutes of Health, Bethesda, Maryland, USA kapilbharti@nei.nih.gov.
authors.forename: Kapil
authors.initials: K
authors.lastname: Bharti
keywords: Cell authentication; Cellular therapy; Genetic differences; Induced pluripotent stem cell (iPSC); Retinal pigment epithelium