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Systematic Comparison of Retinal Organoid Differentiation from Human Pluripotent Stem Cells Reveals Stage Specific, Cell Line, and Methodological Differences

Lookup NU author(s): Dr Carla Jackson, Dr Joseph Collin, Dr Rachel Queen, Dr Birthe HilgenORCiD, Dr Darin Zerti, Majed Felemban, Dr Kathryn White, Professor Evelyne SernagorORCiD, Professor Majlinda LakoORCiD



This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (CC BY-NC-ND).


A major goal in the stem cell field is to generate tissues which can be utilised as a universal tool for in vitro models of development and disease, drug development, or as a resource for patients suffering from disease or injury. Great efforts are being made to differentiate human pluripotent stem cells in vitro towards retinal tissue, which is akin to native human retina in its cytoarchitecture and function, yet the numerous existing retinal induction protocols remain variable in their efficiency and do not routinely produce morphologically or functionally mature photoreceptors. Herein, we determine the impact thatthe method of embryoid body (EB) formation and maintenance as well as cell line background has on retinal organoid differentiation from human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs). Our data indicate that cell line specific differences dominate the variables that underline the differentiation efficiency in the early stages of differentiation. In contrast, the EB generation method and maintenance conditions determine the later differentiation and maturation of retinal organoids. Of the latter, the mechanical method of EB generation under static conditions, accompanied by media supplementation with Y27632 for the first 48 hours of differentiation, results in the most consistent formation of laminated retinal neuroepithelium containing mature and electrophysiologically responsive photoreceptors. Collectively our data provide substantive evidence for stage-specific differences in the ability to give rise to laminated retinae, which is determined by cell line specific differences in the early stages of differentiation and EB generation/maintenance methods at later stages.

Publication metadata

Author(s): Mellough CB, Collin J, Queen R, Hilgen G, Dorgau B, Zerti D, Felemban M, White K, Sernagor E, Lako M

Publication type: Article

Publication status: Published

Journal: STEM CELLS Translational Medicine

Year: 2019

Volume: 8

Issue: 7

Pages: 694-706

Print publication date: 25/06/2019

Online publication date: 27/03/2019

Acceptance date: 11/02/2019

Date deposited: 12/02/2019

ISSN (print): 2157-6564

ISSN (electronic): 2157-6580

Publisher: Wiley


DOI: 10.1002/sctm.18-0267


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Funder referenceFunder name
099175/Z/12/ZWellcome Trust
614620European Research Council (ERC)
MC_PC_15030Medical Research Council (MRC)
GR584RETINA UK (Formerly known as British Retinitis Pigmentosa Society)