Rapid and Efficient Generation of Myelinating Human Oligodendrocytes in Organoids.

Mohammed R Shaker,Ernst J Wolvetang,Ju-Hyun Lee,Sally Martin,Woong Sun,Giovanni Pietrogrande

doi:10.3389/fncel.2021.631548

Mohammed R Shaker, Ernst J Wolvetang + Show 4 more

Open Access

https://doi.org/10.3389/fncel.2021.631548

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Abstract

Human stem cell derived brain organoids are increasingly gaining attention as an ideal model system for investigating neurological diseases, particularly those that involve myelination defects. However, current protocols for generating brain organoids with sufficiently mature oligodendrocytes that deposit myelin on endogenously produced neurons are lengthy and complicated. Taking advantage of a human pluripotent stem cell line that reports on SOX10 expression, we developed a protocol that involves a 42 day exposure of neuroectoderm-derived organoids to a cocktail of growth factors and small molecules that collectively foster oligodendrocyte specification and survival. Importantly, the resulting day 42 brain organoids contain both myelinating oligodendrocytes, cortical neuronal cells and astrocytes. These oligodendrocyte brain organoids therefore constitute a valuable and tractable platform for functional neurogenomics and drug screening for white matter diseases.

Highlights

Oligodendrocytes (OL) are cells of the central nervous system (CNS) that generate the multilayered myelin membrane sheath around vertebrate axons, enhancing the propagation of action potentials
We created a hiPSC line that reports on the expression of the early oligodendroglial gene SOX10 and used this cell line to develop a facile and rapid 42 day protocol for the generation of human brain organoids that contain OL. We show that these OL brain organoids contain OL that myelinate endogenously cospecified cortical neurons and support astrocyte differentiation
From as early as day 3, we exposed the organoids to a cocktail of growth factors and small molecules (Figure 1A), that included thyroid hormone T3, neurotrophin NT3, hepatocytes growth factor (HGF), insulin growth factor (IGF), and platelet derived growth factor (PDGF-AA) to promote differentiation toward the oligodendroglial lineage and to foster the proliferation of newly generated OPCs, as well as B27 without vitamin A and cAMP to drive the maturation of OPCs to myelinating OL (Douvaras and Fossati, 2015; Zhang et al, 2019)

Summary

Introduction

Oligodendrocytes (OL) are cells of the central nervous system (CNS) that generate the multilayered myelin membrane sheath around vertebrate axons, enhancing the propagation of action potentials. OL provide metabolic support to neurons and are important for maintaining axonal integrity (Fünfschilling et al, 2012). OL generation, turnover and dysfunction have been linked to demyelination, axonal damage and disease progression (Domingues et al, 2016). A loss of myelination and a consequent breakdown of OL-axon communication are increasingly linked to neurodegenerative diseases, including amyotrophic lateral sclerosis (Salameh et al, 2015), Parkinson (Ferrer, 2018), and Alzheimer disease (McKenzie et al, 2017). Therapies aimed at modulating OL function and survival, as well as delivery of exogenous healthy OL progenitors (OPCs) are promising avenues for restoring axonal integrity and neurological function

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