Oligodendrogenesis and myelination in a brain microphysiological system
2023
Johns Hopkins University, Baltimore, USA
Oligodendrocytes (OLs), the myelin-forming cells of the central nervous system (CNS), can be differentiated from human induced pluripotent stem cells (hiPSCs), but in vitro modelling of axon myelination in human cells remains challenging. Brain microphysiological systems (bMPS) such as organoids provide an ideal system for studying this process, as OLs differentiate in a more in vivo-like environment, surrounded by neurons and astrocytes that support axon myelination.
Here, the benefits of CRISPR/Cas9 technology were taken advantage of to use reverse transfection to tag the OLs marker proteolipid protein 1 (PLP1) with a fluorescent fusion tag. By differentiating the resulting cell lines in the 3D brain model, the reliability, specificity, and function of the labelled PLP protein were verified and different stages of oligodendrogenesis were followed. Treatment of bMPS with cuprizone resulted in changes in the percentage of labelled cells.
This work demonstrates an efficient method to generate tagged hiPSC lines and the description of a new 3D model to study OL differentiation, migration, and maturation both during in vitro neurodevelopment and in response to environmental chemicals or disease-related stressors.
Oligodendrogenesis and myelination tracing in a CRISPR/Cas9-engineered brain microphysiological system
Lena Smirnova
Added on: 03-14-2023
[1] https://www.frontiersin.org/articles/10.3389/fncel.2022.1094291/full?utm_source=Email_to_authors_&utm_medium=Email&utm_content=T1_11.5e1_author&utm_campaign=Email_publication&field=&journalName=Frontiers_in_Cellular_Neuroscience&id=1094291&emci=c5bf5d33-63a9-ed11-994d-00224832eb73&emdi=470ee51a-75a9-ed11-994d-00224832eb73&ceid=2015591
