Oligodendrocytes (OLs) are responsible for myelin production and metabolic support of neurons. Defects in OLs are
crucial in several neurodegenerative diseases including multiple sclerosis (MS) and amyotrophic lateral sclerosis (ALS).
This protocol describes a method to generate oligodendrocyte precursor cells (OPCs) from human pluripotent stem cells
(hPSCs) in only ~20 d, which can subsequently myelinate neurons, both in vitro and in vivo. To date, OPCs have been
derived from eight different hPSC lines including those derived from patients with spontaneous and familial forms of MS
and ALS, respectively. hPSCs, fated for 8 d toward neural progenitors, are transduced with an inducible lentiviral vector
encoding for SOX10. The addition of doxycycline for 10 d results in >60% of cells being O4-expressing OPCs, of which
20% co-express the mature OL marker myelin basic protein (MBP). The protocol also describes an alternative for viral
transduction, by incorporating an inducible SOX10 in the safe harbor locus AAVS1, yielding ~100% pure OPCs. O4+ OPCs
can be purified and either cryopreserved or used for functional studies. As an example of the type of functional study for
which the derived cells could be used, O4+ cells can be co-cultured with maturing hPSC-derived neurons in 96/384-wellformat plates, allowing the screening of pro-myelinating compounds.
