Abstract
The availability of otherwise not readily accessible intraocular cells via induced pluripotent stem cell (iPSC) technology offers great potential for disease modelling, drug screening, and cell-based transplantation therapy in degenerative ocular disorders. Direct differentiation of iPSCs into retinal pigment epithelium (RPE) is particularly straightforward, and iPSC-derived RPE cell cultures have been demonstrated to yield pure populations of functional cells that display many features of native RPE. Here, I describe a protocol for the generation of iPSC-derived RPE monolayer, their propagation, and cryostorage. A reliable monitoring for functional cell differentiation is achieved by measuring transepithelial resistance.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Takahashi K, Tanabe K, Ohnuki M et al (2007) Induction of pluripotent stem cells from adult human fibroblasts by defined factors. Cell 131:861–872
Takahashi K, Yamanaka S (2016) A decade of transcription factor-mediated reprogramming to pluripotency. Nat Rev Mol Cell Biol 17:183–193
Ramsden CM, Powner MB, Carr AJ et al (2013) Stem cells in retinal regeneration: past, present and future. Development 140:2576–2585
Brandl C, Zimmermann SJ, Milenkovic VM et al (2014) In-depth characterisation of retinal pigment epithelium (RPE) cells derived from human induced pluripotent stem cells (hiPSC). NeuroMolecular Med 16:551–564
Buchholz DE, Pennington BO, Croze RH et al (2013) Rapid and efficient directed differentiation of human pluripotent stem cells into retinal pigmented epithelium. Stem Cells Transl Med 2:384–393
Du H, Lim SL, Grob S et al (2011) Induced pluripotent stem cell therapies for geographic atrophy of age-related macular degeneration. Semin Ophthalmol 26:216–224
Kamao H, Mandai M, Okamoto S et al (2014) Characterization of human induced pluripotent stem cell-derived retinal pigment epithelium cell sheets aiming for clinical application. Stem Cell Rep 2:205–218
Krohne TU, Westenskow PD, Kurihara T et al (2012) Generation of retinal pigment epithelial cells from small molecules and OCT4 reprogrammed human induced pluripotent stem cells. Stem Cells Transl Med 1:96–109
Singh R, Phillips MJ, Kuai D et al (2013) Functional analysis of serially expanded human iPS cell-derived RPE cultures. Invest Ophthalmol Vis Sci 54:6767–6778
Ablonczy Z, Dahrouj M, Tang PH et al (2011) Human retinal pigment epithelium cells as functional models for the RPE in vivo. Invest Ophthalmol Vis Sci 52:8614–8620
Acknowledgments
I gratefully acknowledge the excellent and dedicated cell culture work of Nico Hertel. This study was in part supported by a grant from Novartis Pharma GmbH, Nürnberg, Germany (Reference No. 3625340) and Institutional Support from Titel 77 (Institute of Human Genetics, University of Regensburg).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Brandl, C. (2019). Generation of Functional Retinal Pigment Epithelium from Human Induced Pluripotent Stem Cells. In: Weber, B.H.F., Langmann, T. (eds) Retinal Degeneration. Methods in Molecular Biology, vol 1834. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-8669-9_6
Download citation
DOI: https://doi.org/10.1007/978-1-4939-8669-9_6
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-4939-8668-2
Online ISBN: 978-1-4939-8669-9
eBook Packages: Springer Protocols