Differentiation of brain and retinal organoids from confluent cultures of pluripotent stem cells connected by nerve-like axonal projections of optic origin

Milan Fernando; Scott Lee; Jesse R Wark; Di Xiao; Benjamin Y Lim; Michelle O'Hara-Wright; Hani J Kim; Grady C Smith; Ted Wong; Erdahl T Teber; Robin R Ali; Pengyi Yang; Mark E Graham; Anai Gonzalez-Cordero

doi:10.1016/j.stemcr.2022.04.003

Differentiation of brain and retinal organoids from confluent cultures of pluripotent stem cells connected by nerve-like axonal projections of optic origin

Stem Cell Reports. 2022 Jun 14;17(6):1476-1492. doi: 10.1016/j.stemcr.2022.04.003. Epub 2022 May 5.

Authors

Affiliations

¹ Stem Cell Medicine and Stem Cell and Organoid Facility, University of Sydney, Westmead, 2145 NSW, Australia.
² Stem Cell Medicine and Stem Cell and Organoid Facility, University of Sydney, Westmead, 2145 NSW, Australia; Synapse Proteomics, University of Sydney, Westmead, 2145 NSW, Australia.
³ Computational Systems Biology, University of Sydney, Westmead, 2145 NSW, Australia; Charles Perkins Centre, School of Mathematics and Statistics, University of Sydney, Sydney, NSW 2006, Australia; School of Medical Sciences, Faculty of Medicine and Health, University of Sydney, NSW 2006, Australia.
⁴ Stem Cell Medicine and Stem Cell and Organoid Facility, University of Sydney, Westmead, 2145 NSW, Australia; School of Medical Sciences, Faculty of Medicine and Health, University of Sydney, NSW 2006, Australia.
⁵ Bioinformatics, Children's Medical Research Institute, University of Sydney, Westmead, 2145 NSW, Australia.
⁶ Bioinformatics, Children's Medical Research Institute, University of Sydney, Westmead, 2145 NSW, Australia; School of Medical Sciences, Faculty of Medicine and Health, University of Sydney, NSW 2006, Australia.
⁷ Gene and Cell Therapy Group, UCL Institute of Ophthalmology, London EC1V 9EL, UK.
⁸ Synapse Proteomics, University of Sydney, Westmead, 2145 NSW, Australia; School of Medical Sciences, Faculty of Medicine and Health, University of Sydney, NSW 2006, Australia.
⁹ Stem Cell Medicine and Stem Cell and Organoid Facility, University of Sydney, Westmead, 2145 NSW, Australia; School of Medical Sciences, Faculty of Medicine and Health, University of Sydney, NSW 2006, Australia. Electronic address: agonzalez-cordero@cmri.org.au.

Abstract

Advances in the study of neurological conditions have been possible because of pluripotent stem cell technologies and organoids. Studies have described the generation of neural ectoderm-derived retinal and brain structures from pluripotent stem cells. However, the field is still troubled by technical challenges, including high culture costs and variability. Here, we describe a simple and economical protocol that reproducibly gives rise to the neural retina and cortical brain regions from confluent cultures of stem cells. The spontaneously generated cortical organoids are transcriptionally comparable with organoids generated by other methods. Furthermore, these organoids showed spontaneous functional network activity and proteomic analysis confirmed organoids maturity. The generation of retinal and brain organoids in close proximity enabled their mutual isolation. Suspension culture of this complex organoid system demonstrated the formation of nerve-like structures connecting retinal and brain organoids, which might facilitate the investigation of neurological diseases of the eye and brain.

Keywords: brain organoids; multielectrode array; omics; optic axonal projections; pluripotent stem cells; retinal organoids.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Brain
Cell Differentiation
Induced Pluripotent Stem Cells*
Organoids
Pluripotent Stem Cells*
Proteomics
Retina