RT Journal Article
T1 Generation of oligodendrocytes and establishment of an all-human myelinating platform from human pluripotent stem cells
A1 García-León, Juan Antonio
A1 Garcia Diaz, Beatriz
A1 Eggermont, Kristel
A1 Cáceres Palomo, Laura
A1 Neyrinck, Katrien
A1 Madeiro da Costa, Rodrigo
A1 Dávila-Cansino, José Carlos
A1 Baron Van Evercooren, Anne
A1 Gutiérrez-Pérez, Antonia
A1 Verfaille, Catherine M
K1 Neuronas
AB Oligodendrocytes (OLs) are responsible for myelin production and metabolic support of neurons. Defects in OLs arecrucial 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 beenderived from eight different hPSC lines including those derived from patients with spontaneous and familial forms of MSand ALS, respectively. hPSCs, fated for 8 d toward neural progenitors, are transduced with an inducible lentiviral vectorencoding for SOX10. The addition of doxycycline for 10 d results in >60% of cells being O4-expressing OPCs, of which20% co-express the mature OL marker myelin basic protein (MBP). The protocol also describes an alternative for viraltransduction, by incorporating an inducible SOX10 in the safe harbor locus AAVS1, yielding ~100% pure OPCs. O4+ OPCscan be purified and either cryopreserved or used for functional studies. As an example of the type of functional study forwhich 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.
PB Springer Nature
YR 2020
FD 2020
LK https://hdl.handle.net/10630/32616
UL https://hdl.handle.net/10630/32616
LA eng
NO García-León, J.A., García-Díaz, B., Eggermont, K. et al. Generation of oligodendrocytes and establishment of an all-human myelinating platform from human pluripotent stem cells. Nat Protoc 15, 3716–3744 (2020). https://doi.org/10.1038/s41596-020-0395-4
DS RIUMA. Repositorio Institucional de la Universidad de Málaga
RD 21 ene 2026