Human Myelin Spheres for In Vitro Oligodendrocyte Maturation, Myelination and Neurological Disease Modeling

We used single nucleus RNA sequencing (10x genomics) to characterize a novel 3D spheroid model - Myelin Spheres (MyS). Neuron Monoculture Spheroids (NMS) as controls. MyS comprises of neuronal progenitor cells (NPCs) with oligodendrocyte progenitor cells (OPCs) while NMS were made of only NPCs. Both NPCs and OPCs were derived from human induced pluripotent stem cells (hiPSCs) with or without an inducible SOX10 casette integrated in the genome. We performed sequencing on MyS and NMS at week 8 and 15 respectively, to understand the temporal change in oligodendrocyte maturation. Overall design: We used four to six spheroids for both MyS or NMS as one sample, for single nucleus extraction on week 8 and 15 seperately. Extracted nuclei were counted and checked for quality and were then used for library preparation and sequencing.