Figure 3. Human GPCs differentiate as myelinogenic oligodendrocytes in response to cuprizone demyelination.

A, Mice were transplanted with 2×10⁵ hGPCs perinatally, and at 17 wks of age placed on either a cuprizone (CZN)-supplemented or normal diet for 12 wks, then either sacrificed or returned to standard diet and killed at later time-points. B–C. Serial coronal sections comparing dot-mapped distributions of human (human nuclear antigen, hN) cells in control (B) and cuprizone-fed mice at 49 wks of age, after 20 weeks recovery on control diet. D–E. Relative abundance of human (red dots) and mouse (blue) transferrin (TF)-defined oligodendroglia, in 20 μm coronal sections of corpus callosa of mice engrafted with hGPCs neonatally, demyelinated as adults from 17–29 wks of age, then assessed at 49 wks, 20 wks after cuprizone. E shows an untreated control, age-matched to D. F. The density of human cells in the corpus callosum increases to a greater degree in cuprizone-demyelinated brains than in untreated controls, including during the period of cuprizone treatment, indicating progenitor mobilization. G, By 8 weeks after the termination of cuprizone exposure, the density of human oligodendroglia is >5-fold greater in cuprizone-demyelinated than untreated control brains. H, By that 8-week recovery point, most hGPCs engrafted in the corpus callosa of cuprizone-treated mice differentiated as oligodendrocytes, and accordingly (I), over half of all transferrin-defined oligodendrocytes were human; in contrast, relatively few human oligodendrocytes were noted in untreated chimeric brains. J, Substantial colonization by human glia is evident in this remyelinated callosum, after a 20-week recovery (human nuclear antigen, magenta; myelin basic protein, green). K, chimeric white matter populated, post- cuprizone, by human GPC-derived oligodendroglia. Anti-human nuclear antigen (hNA) (red), transferrin, (green); inset shows relative abundance of hNA⁺/transferrin⁺ human oligodendroglia. Scale: J, 100 μm; K, 50 μm, inset, 25 μm.

From (Windrem et al., 2020).