Fast generation of forebrain oligodendrocyte spheroids from human embryonic stem cells by transcription factors

Oligodendrocyte spheroids (OL-spheroids) containing oligodendrocytes and neurons provide an accessible system to dissect demyelinating diseases and test therapeutic treatment. However, generation of human OL-spheroids is still technically challenging and time-consuming until now. Here, we presented evidence that overexpression of SOX10 and OLIG2 (SO) in human embryonic stem cells (hESCs)-derived ventral forebrain neural progenitors is sufficient to produce forebrain pre-oligodendrocytes (pre-OLs) and mature oligodendrocytes (OLs) within 20-40 days. More importantly, optimizing this procedure by overexpression of SO in ventral forebrain spheroids, we successfully generated OL-spheroids with pre-OLs, matureOLs, and neurons 40 days after OL-induction. We further demonstrated oligodendrocyte-neuron interactions and obvious axonmyelination inOLspheroids. Finally, over 30% cells developed into mature oligodendrocytes with forebrain identity and myelinate axons in mouse brain 3 months after transplantation. This study provides a strategy to generate forebrain OL-spheroids rapidly and efficiently which would facilitate development of new therapeutics for demyelinating disorders.

Automated summary

This study presents a strategy to rapidly and efficiently generate forebrain oligodendrocyte spheroids by overexpressing SOX10 and OLIG2 in human embryonic stem cells. The generated spheroids contain mature oligodendrocytes, pre-oligodendrocytes, and neurons, providing a valuable system for studying demyelinating diseases and testing therapeutic treatments. Transplantation of these spheroids into mouse brains resulted in the development of mature oligodendrocytes that myelinated axons, demonstrating the potential for this method in developing new therapeutics.