Human embryonic stem cells differentiation into oligodendrocyte lineage cells v1

Oligodendrocyte (OL) lineage cell generation from human embryonic stem cell line Royan H6 (RH6) started by dual inhibition of SMAD signaling. Therefore, within eight days, adherent NANOG+RH6 cells (B) differentiated into SOX1+ neural stem cells (C). Next, the embryonic ventral spinal cord environment (pMN domain) was mimicked by the application of two morphogens, RA and SAG and the OLIG2+ neural progenitor cells were generated on day 12 (D) of differentiation. Further, oligodendrocyte progenitor cell commitment was achieved on day 20, when the pre-oligodendrocyte progenitor cell (pre-OPC) aggregates were positively stained by NKX2.2+ (E). From day 20 to day 80, supportive reagents for pre-OPCs maturation into PDGFRA+ OPCs (F), and their further expansion and maturation toward oligodendrocyte producing OPCs were added to the culture medium. Finally, on day 80 the growth-factors were withdrawn from the culture medium and OPCs were differentiated into MBP+ OLs (G). References: - Baharvand H, Ashtiani SK, Taee A, Massumi M, Valojerdi MR, Yazdi PE, et al. Generation of new human embryonic stem cell lines with diploid and triploid karyotypes. Dev Growth Differ. 2006;48:117–28. - Douvaras P, Wang J, Zimmer M, Hanchuk S, O’Bara MA, Sadiq S, et al. Efficient generation of myelinating oligodendrocytes from primary progressive multiple sclerosis patients by induced pluripotent stem cells. Stem Cell Reports [Internet]. The Authors; 2014;3:250–9. Available from: http://dx.doi.org/10.1016/j.stemcr.2014.06.012 - Douvaras P, Fossati V. Generation and isolation of oligodendrocyte progenitor cells from human pluripotent stem cells. Nat Protoc [Internet]. 2015;10:1143–54. Available from: http://dx.doi.org/10.1038/nprot.2015.075 - Piao J, Major T, Auyeung G, Policarpio E, Menon J, Droms L, et al. Human embryonic stem cell-derived oligodendrocyte progenitors remyelinate the brain and rescue behavioral deficits following radiation. Cell Stem Cell [Internet]. Elsevier Inc.; 2015;16:198–210. Available from: http://dx.doi.org/10.1016/j.stem.2015.01.004