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oai:riuma.uma.es:10630/326162026-02-03T11:29:44Zcom_10630_2254col_10630_37953

García-León, Juan Antonio Garcia Diaz, Beatriz Eggermont, Kristel Cáceres Palomo, Laura Neyrinck, Katrien Madeiro da Costa, Rodrigo Dávila-Cansino, José Carlos Baron Van Evercooren, Anne Gutiérrez-Pérez, Antonia Verfaille, Catherine M 2024-09-18T10:56:43Z 2024-09-18T10:56:43Z 2020 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 https://hdl.handle.net/10630/32616 10.1038/s41596-020-0395-4 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. eng open access Neuronas Generation of oligodendrocytes and establishment of an all-human myelinating platform from human pluripotent stem cells journal article