@article{c24f9b13b6944d36a6c47bee2d97d3ce,
title = "Cell Types of the Human Retina and Its Organoids at Single-Cell Resolution",
abstract = "Human organoids recapitulating the cell-type diversity and function of their target organ are valuable for basic and translational research. We developed light-sensitive human retinal organoids with multiple nuclear and synaptic layers and functional synapses. We sequenced the RNA of 285,441 single cells from these organoids at seven developmental time points and from the periphery, fovea, pigment epithelium and choroid of light-responsive adult human retinas, and performed histochemistry. Cell types in organoids matured in vitro to a stable “developed” state at a rate similar to human retina development in vivo. Transcriptomes of organoid cell types converged toward the transcriptomes of adult peripheral retinal cell types. Expression of disease-associated genes was cell-type-specific in adult retina, and cell-type specificity was retained in organoids. We implicate unexpected cell types in diseases such as macular degeneration. This resource identifies cellular targets for studying disease mechanisms in organoids and for targeted repair in human retinas.",
keywords = "eye disease, human retina, macular degeneration, organoid, organoid development, retina, retinal organoid, single cell sequencing, synaptic function, transcriptome",
author = "Cowan, {Cameron S.} and Magdalena Renner and {De Gennaro}, Martina and Brigitte Gross-Scherf and David Goldblum and Yanyan Hou and Martin Munz and Rodrigues, {Tiago M.} and Jacek Krol and Tamas Szikra and Rachel Cuttat and Annick Waldt and Panagiotis Papasaikas and Roland Diggelmann and Patino-Alvarez, {Claudia P.} and Patricia Galliker and Spirig, {Stefan E.} and Dinko Pavlinic and Nadine Gerber-Hollbach and Sven Schuierer and Aldin Srdanovic and Marton Balogh and Riccardo Panero and Akos Kusnyerik and Arnold Szabo and Stadler, {Michael B.} and Selim Org{\"u}l and Simone Picelli and Hasler, {Pascal W.} and Andreas Hierlemann and Scholl, {Hendrik P.N.} and Guglielmo Roma and Florian Nigsch and Botond Roska",
note = "Funding Information: We would like to thank the organ and tissue donors and their families for their generous contributions to science; T. V{\"o}gele, J. Sprachta, and P. Blaschke the transplant coordinators at the University Hospital Basel for organizing organ donations; Z. Raics and D. Hillier for developing recording software; P. Argast and P. Buchmann for electrical and mechanical engineering in support of the experiments; P. Ala-Laurila for advice on primate retinal electrophysiology; S. Rompani for advice on organoid physiology; W. Baehr and A. Neutzner for sharing of antibodies. We thank IOB, FMI, and NIBR core facilities for their support, especially the FMI microscopy and imaging facility, in particular C. Genoud and A. Graff-Meyer for electron microscopy sample preparation and image acquisition. We thank the group of G. Roma for support with scRNA sequencing. We thank the IOB complex virus platform for providing GCaMP6s AAV for organoid physiology experiments. We also thank P. King, F. Franke, J. Matsell, and E. Sigle for commenting on the manuscript. We thank the members of the Roska, Nigsch, Roma, and Hierlemann laboratories for discussions on the manuscript. This work was supported by the following grants: Swiss National Science Foundation Synergia ( CRSII3_141801 to A.H. and B.R.), European Research Council Advanced Grant (to B.R.), Gebert-R{\"u}f (to B.R.), Swiss National Science Foundation (to B.R. and H.P.N.S.), NCCR “Molecular Systems Engineering” (to B.R. and H.P.N.S.), Lynn and Diana Lady Dougan private donation (to B.R.), Wellcome Trust (to H.P.N.S.), and Foundation Fighting Blindness (to H.P.N.S.). Funding Information: We would like to thank the organ and tissue donors and their families for their generous contributions to science; T. V{\"o}gele, J. Sprachta, and P. Blaschke the transplant coordinators at the University Hospital Basel for organizing organ donations; Z. Raics and D. Hillier for developing recording software; P. Argast and P. Buchmann for electrical and mechanical engineering in support of the experiments; P. Ala-Laurila for advice on primate retinal electrophysiology; S. Rompani for advice on organoid physiology; W. Baehr and A. Neutzner for sharing of antibodies. We thank IOB, FMI, and NIBR core facilities for their support, especially the FMI microscopy and imaging facility, in particular C. Genoud and A. Graff-Meyer for electron microscopy sample preparation and image acquisition. We thank the group of G. Roma for support with scRNA sequencing. We thank the IOB complex virus platform for providing GCaMP6s AAV for organoid physiology experiments. We also thank P. King, F. Franke, J. Matsell, and E. Sigle for commenting on the manuscript. We thank the members of the Roska, Nigsch, Roma, and Hierlemann laboratories for discussions on the manuscript. This work was supported by the following grants: Swiss National Science Foundation Synergia (CRSII3_141801 to A.H. and B.R.), European Research Council Advanced Grant (to B.R.), Gebert-R{\"u}f (to B.R.), Swiss National Science Foundation (to B.R. and H.P.N.S.), NCCR “Molecular Systems Engineering” (to B.R. and H.P.N.S.), Lynn and Diana Lady Dougan private donation (to B.R.), Wellcome Trust (to H.P.N.S.), and Foundation Fighting Blindness (to H.P.N.S.). M.B.S. S.O. S.P. A.H. H.P.N.S. G.R. F.N. and B.R. designed and supervised experiments. M.R. B.G.-S. Y.H. M.M. J.K. C.P.P.-A. P.G. and A. Srdanovic coordinated, optimized, and performed iPSC and organoid generation. C.S.C. M.R. B.G.-S. T.M.R. R.C. A.W. D.P. S.P. and G.R. coordinated, optimized, and performed sequencing experiments. C.S.C. D.G. T.S. N.G.-H. M.B. A.K. A. Szabo, and P.H. coordinated, optimized, and performed retinal donations. S.E.S. optimized AAV infection of organoids. C.S.C. R.D. and M.B. optimized and performed retinal electrophysiology experiments. C.S.C. M.D.G. and M.M. performed organoid calcium imaging experiments. M.R. B.G.-S. Y.H. M.M. and T.M.R. optimized and performed retinal and organoid immunostaining, in situ hybridization, and confocal imaging. C.S.C. P.P. and S.S. performed analysis of sequencing data. C.S.C. and R.D. performed analysis of retinal and organoid physiology data. C.S.C. M.R. T.M.R. and B.R. designed and performed statistical analyses. R.P. performed development of tools to visualize sequencing data. C.S.C. M.R. and B.R. wrote the paper. The authors declare no competing interests. Publisher Copyright: {\textcopyright} 2020 The Author(s)",
year = "2020",
month = sep,
day = "17",
doi = "10.1016/j.cell.2020.08.013",
language = "English",
volume = "182",
pages = "1623--1640.e34",
journal = "Cell",
issn = "0092-8674",
publisher = "Cell Press",
number = "6",
}