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

Carla B. Mellough, Joseph Collin, Rachel Queen, Gerrit Hilgen, Birthe Dorgau, Darin Zerti, Majed Felemban, Kathryn White, Evelyne Sernagor, Majlinda Lako

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

A major goal in the stem cell field is to generate tissues which can be utilized 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 toward 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 that the method of embryoid body (EB) formation and maintenance as well as cell line background has on retinal organoid differentiation from human embryonic stem cells and human induced pluripotent stem cells. 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/organoid maintenance methods at later stages.

Original languageEnglish
Pages (from-to)694-706
JournalStem Cells Translational Medicine
Volume8
Issue number7
DOIs
Publication statusPublished - Jul 2019

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Organoids
Embryoid Bodies
Pluripotent Stem Cells
Cell Line
Maintenance
Retina
Induced Pluripotent Stem Cells
Population Groups
Stem Cells
Wounds and Injuries
Pharmaceutical Preparations
In Vitro Techniques

Cite this

Mellough, Carla B. ; Collin, Joseph ; Queen, Rachel ; Hilgen, Gerrit ; Dorgau, Birthe ; Zerti, Darin ; Felemban, Majed ; White, Kathryn ; Sernagor, Evelyne ; Lako, Majlinda. / Systematic Comparison of Retinal Organoid Differentiation from Human Pluripotent Stem Cells Reveals Stage Specific, Cell Line, and Methodological Differences. In: Stem Cells Translational Medicine. 2019 ; Vol. 8, No. 7. pp. 694-706.
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Systematic Comparison of Retinal Organoid Differentiation from Human Pluripotent Stem Cells Reveals Stage Specific, Cell Line, and Methodological Differences. / Mellough, Carla B.; Collin, Joseph; Queen, Rachel; Hilgen, Gerrit; Dorgau, Birthe; Zerti, Darin; Felemban, Majed; White, Kathryn; Sernagor, Evelyne; Lako, Majlinda.

In: Stem Cells Translational Medicine, Vol. 8, No. 7, 07.2019, p. 694-706.

Research output: Contribution to journalArticle

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AU - Hilgen, Gerrit

AU - Dorgau, Birthe

AU - Zerti, Darin

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AU - Lako, Majlinda

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AB - A major goal in the stem cell field is to generate tissues which can be utilized 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 toward 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 that the method of embryoid body (EB) formation and maintenance as well as cell line background has on retinal organoid differentiation from human embryonic stem cells and human induced pluripotent stem cells. 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/organoid maintenance methods at later stages.

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