Title:

Promoting Oligodendrocyte Differentiation from Human Neural Stem Cells for 3D Myelination Models

Oligodendrocyte-mediated myelination is essential for efficient neuronal signaling and is disrupted in neurological disorders such as multiple sclerosis. In this study, we evaluated a gelatin-based microgel system as a three-dimensional (3D) platform to promote oligodendrocyte differentiation from human neural stem cells (ReNcell VM), compared to traditional two-dimensional (2D) culture. Cells were cultured under growth and oligodendrocyte differentiation conditions and assessed using microscopy and immunofluorescence for lineage markers. Across all conditions, minimal expression of oligodendrocyte markers (Olig2, MBP) was observed, indicating limited differentiation and myelin formation. Both 2D and 3D systems showed similar outcomes, suggesting that current media conditions, rather than culture dimensionality, limit differentiation. These findings highlight the need for optimized media composition and extended differentiation timelines to support effective oligodendrocyte maturation in vitro.

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