A method of differentiating
nonhuman primate induced pluripotent stem cells (iPSCs) to oligodendrocytes is provided. The method may include providing a modified vector comprising a piggyBac vector backbone that expresses one or more differentiation factor genes, including SOX10, OLIG2, and NKX6-2. The piggyBac vector backbone may comprise one or more terminal inverted repeats and one or more
transposase recognition sites configured to interact with a piggyBac
transposase to mediate genomic integration at TTAA or noncanonical target sequences. The vector may further include a
selectable marker and a
promoter for regulated expression. The modified vector may be introduced into
nonhuman primate iPSCs, for example from Macaca fascicularis, by
electroporation and used in a feeder-free differentiation
workflow. The resulting differentiated cells may be identified by expression of
oligodendrocyte-associated markers including O4, O1,
PDGFRA, MOG, GALC, CNPase, and
myelin basic
protein (MBP).