Protocol for generating embedding-free brain organoids enriched with oligodendrocytes

Bahaa Al-Mhanawi; Marta Boira Marti; Sean D Morrison; Pallavi Gupta; Maath Alani; Peter G Noakes; Ernst J Wolvetang; Mohammed R Shaker

doi:10.1016/j.xpro.2023.102725

Protocol for generating embedding-free brain organoids enriched with oligodendrocytes

STAR Protoc. 2023 Dec 15;4(4):102725. doi: 10.1016/j.xpro.2023.102725. Epub 2023 Nov 16.

Authors

Bahaa Al-Mhanawi¹, Marta Boira Marti¹, Sean D Morrison¹, Pallavi Gupta², Maath Alani¹, Peter G Noakes³, Ernst J Wolvetang⁴, Mohammed R Shaker⁵

Affiliations

¹ Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St. Lucia, QLD 4072, Australia.
² Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St. Lucia, QLD 4072, Australia; University of Queensland - IIT Delhi Academy of Research (UQIDAR), Hauz Khas, New Delhi 110016, India; Department of Biochemical Engineering and Biotechnology, Indian Institute of Technology Delhi, Hauz Khas, New Delhi 110016, India.
³ Queensland Brain Institute, The University of Queensland, St. Lucia, QLD 4072, Australia; School of Biomedical Sciences, The University of Queensland, St. Lucia, QLD 4072, Australia.
⁴ Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St. Lucia, QLD 4072, Australia. Electronic address: e.wolvetang@uq.edu.au.
⁵ Australian Institute for Bioengineering and Nanotechnology, The University of Queensland, St. Lucia, QLD 4072, Australia. Electronic address: m.shaker@uq.edu.au.

Abstract

In response to the scarcity of advanced in vitro models dedicated to human CNS white matter research, we present a protocol to generate neuroectoderm-derived embedding-free human brain organoids enriched with oligodendrocytes. We describe steps for neuroectoderm differentiation, development of neural spheroids, and their transferal to Matrigel. We then detail procedures for the development, maturation, and application of oligodendrocyte-enriched brain organoids. The presence of myelin-producing cells makes these organoids useful for studying human white matter diseases, such as leukodystrophy.

Keywords: Neuroscience; Organoids; Stem Cells.

MeSH terms

Brain*
Humans
Myelin Sheath
Oligodendroglia*
Organoids