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Human induced pluripotent stem cells generate light responsive retinal organoids with variable and nutrient dependent efficiency

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posted on 29.01.2021, 11:28 authored by Dean Hallam, Gerrit Hilgen, Birthe Dorgau, Lili Zhu, Min Yu, Sanja Bojic, Philip Hewitt, Michael Schmitt, Marianne Uteng, Stefan Kustermann, David Steel, Mike Nicholds, Rob ThomasRob Thomas, Achim Treumann, Andrew Porter, Evelyne Sernagor, Lyle Armstrong, Majlinda Lako
The availability of in vitro models of the human retina in which to perform pharmacological and toxicological studies is an urgent and unmet need. An essential step for developing in vitro models of human retina is the ability to generate laminated, physiologically functional and light-responsive retinal organoids from renewable and patient specific sources. We investigated five different human induced pluripotent stem cell (iPSC) lines and showed a significant variability in their efficiency to generate retinal organoids. Despite this variability, by month 5 of differentiation, all iPSC-derived retinal organoids were able to generate light responses, albeit immature, comparable to the earliest light responses recorded from the neonatal mouse retina, close to the period of eye opening. All iPSC-derived retinal organoids exhibited at this time a well-formed outer nuclear like layer containing photoreceptors with inner segments, connecting cilium and outer like segments. The differentiation process was highly dependent on seeding cell density and nutrient availability determined by factorial experimental design. We adopted the differentiation protocol to a multiwell plate format which enhanced generation of retinal organoids with retinal pigmented epithelium (RPE) and improved ganglion cell development and the response to physiological stimuli. We tested the response of iPSC-derived retinal organoids to Moxifloxacin and showed that similarly to in vivo adult mouse retina, the primary affected cell types were photoreceptors. Together our data indicate that light responsive retinal organoids derived from carefully selected and differentiation efficient iPSC lines can be generated at the scale needed for pharmacology and drug screening purposes. © AlphaMed Press 2018.

Funding

This work was funded by the European Research Council (#614620], CRACKIT23 challenge phase 1 award (NC/CO16206/1), MRC Confidence in Concept award (MC_PC_15030) and RP Fighting Blindness Innovation grant (GR584).

History

School

  • Mechanical, Electrical and Manufacturing Engineering

Published in

Stem Cells

Volume

36

Issue

10

Pages

1535-1551

Citation

HALLAM, D, ... et al., 2018. Human induced pluripotent stem cells generate light responsive retinal organoids with variable and nutrient dependent efficiency. Stem Cells, 36(10), pp. 1535-1551.

Publisher

© AlphaMed Press 2018

Version

VoR (Version of Record)

Publisher statement

This work is made available according to the conditions of the Creative Commons Attribution 4.0 Unported Licence (CC BY). Full details of this licence are available at: http://creativecommons.org/licenses/by/4.0/

Acceptance date

25/06/2018

Publication date

2018-07-13

Copyright date

2018

Notes

This is an Open Access Article. It is published by AlphaMed Press under the Creative Commons Attribution 4.0 Unported Licence (CC BY). Full details of this licence are available at: http://creativecommons.org/licenses/by/4.0/

eISSN

1549-4918

Language

en

Location

United States