Enhancing Stem Cell Differentiation with a Transient DMSO Treatment

Begin with a multi-well plate containing adhered human pluripotent stem cells with multilineage differentiation ability. 

Introduce a stem cell medium containing dimethyl sulfoxide or DMSO.

DMSO enters the cells and prevents the progression of the cell cycle from the Gap 1, or G1 phase, an initial stage of the cell cycle, to the synthesis, or S phase.

This creates a uniform population of cells in the G1 phase, enhancing their differentiation ability.

Replace the medium with a nutrient-rich ectodermal differentiation medium containing inhibitors of the BMP and TGF-β signaling pathways.

These inhibitors bind to specific ALK receptors on the stem cells, blocking the BMP and TGF-β signaling pathways and preventing their differentiation into endoderm and mesoderm precursors.

This allows the selective differentiation of G1-arrested stem cells into ectodermal precursors possessing neural differentiation ability.

Regularly replace the medium with a fresh medium to maintain the nutrient and differentiation factor levels, ensuring cell survival.