Basic Study
Copyright ©The Author(s) 2020. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Stem Cells. Oct 26, 2020; 12(10): 1171-1183
Published online Oct 26, 2020. doi: 10.4252/wjsc.v12.i10.1171
Enrichment of retinal ganglion and Müller glia progenitors from retinal organoids derived from human induced pluripotent stem cells - possibilities and current limitations
Kristine Karla Freude, Sarkis Saruhanian, Alanna McCauley, Colton Paterson, Madeleine Odette, Annika Oostenink, Poul Hyttel, Mark Gillies, Henriette Haukedal, Miriam Kolko
Kristine Karla Freude, Alanna McCauley, Colton Paterson, Madeleine Odette, Annika Oostenink, Poul Hyttel, Henriette Haukedal, Department of Veterinary and Animal Sciences, Group of Stem Cell Models for Studies of Neurodegenerative Diseases, Section for Pathobiological Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Frederiksberg 1870, Denmark
Sarkis Saruhanian, Miriam Kolko, Department of Drug Design and Pharmacology, University of Copenhagen, Copenhagen 2100, Denmark
Mark Gillies, Save Sight Institute, South Block, Sydney Eye Hospital, Sydney 2000, Australia
Miriam Kolko, Department of Ophthalmology, Copenhagen University Hospital, Rigshospitalet-Glostrup, Glostrup 2600, Denmark
Author contributions: Freude KK and Kolko M designed and coordinated the study; Saruhanian S, McCauley A, Paterson C, Odette M, Oostenink A, Freude KK, and Hyttel P performed the experiments, acquired and analyzed the data; Saruhanian S, McCauley A, Paterson C, Odette M, Hyttel P, Haukedal H, and Freude KK interpreted the data; Gillies M provided intellectual input into the experimental design; Freude KK, Haukedal H and Kolko M wrote the manuscript; all authors approved the final version of the manuscript.
Supported by Innovation Fund Denmark, No. 4108-00008B; The Bagenkop Nielsens Ø jen-Fond, No. 115227; Hø rslev-Fonden, No. 116967; Beckett Fonden, No. 116936; and Velux Foundation, No. 1179261001/2.
Institutional review board statement: The study was reviewed and approved by the Institutional Review Board at the Department of Drug Design and Pharmacology at the University of Copenhagen.
Conflict-of-interest statement: The authors declare they do not have conflicts of interest regarding the publication of this article.
Data sharing statement: No additional data are available.
ARRIVE guidelines statement: The ARRIVE Guidelines have been adopted.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Corresponding author: Kristine Karla Freude, BSc, DPhil, MSc, PhD, Associate Professor, Department of Veterinary and Animal Sciences, Group of Stem Cell Models for Studies of Neurodegenerative Diseases, Section for Pathobiological Sciences, Faculty of Health and Medical Sciences, University of Copenhagen, Grønnegårdsvej 7, Frederiksberg 1870, Denmark. kkf@sund.ku.dk
Received: June 11, 2020
Peer-review started: June 11, 2020
First decision: June 20, 2020
Revised: July 3, 2020
Accepted: August 16, 2020
Article in press: August 16, 2020
Published online: October 26, 2020
Processing time: 137 Days and 2.3 Hours
Abstract
BACKGROUND

Retinal organoids serve as excellent human-specific disease models for conditions affecting otherwise inaccessible retinal tissue from patients. They permit the isolation of key cell types affected in various eye diseases including retinal ganglion cells (RGCs) and Müller glia.

AIM

To refine human-induced pluripotent stem cells (hiPSCs) differentiated into three-dimensional (3D) retinal organoids to generate sufficient numbers of RGCs and Müller glia progenitors for downstream analyses.

METHODS

In this study we described, evaluated, and refined methods with which to generate Müller glia and RGC progenitors, isolated them via magnetic-activated cell sorting, and assessed their lineage stability after prolonged 2D culture. Putative progenitor populations were characterized via quantitative PCR and immunocytochemistry, and the ultrastructural composition of retinal organoid cells was investigated.

RESULTS

Our study confirms the feasibility of generating marker-characterized Müller glia and RGC progenitors within retinal organoids. Such retinal organoids can be dissociated and the Müller glia and RGC progenitor-like cells isolated via magnetic-activated cell sorting and propagated as monolayers.

CONCLUSION

Enrichment of Müller glia and RGC progenitors from retinal organoids is a feasible method with which to study cell type-specific disease phenotypes and to potentially generate specific retinal populations for cell replacement therapies.

Keywords: Human induced pluripotent stem cells; Retinal organoids; Retinal ganglion cells; Müller glia; Progenitors; Cell-type enrichment

Core Tip: Retinal organoids derived from human-induced pluripotent stem cells are excellent tools for enriching specific subpopulations for subsequent studies of cell type-specific disease phenotypes affecting the eye. Here, we describe the generation of retinal organoids and the harvest as well as 2D maintenance of retinal ganglion cells and Müller glia progenitors.