Differentiation of Spinal Cord Neural Progenitor Cells into Motor Neurons

Begin with a culture plate coated with a synthetic polymer and an adhesion protein. 

Add spinal cord neural progenitor cells resuspended in a motor neuron differentiation medium, supplemented with a small-molecule inhibitor.

The coated plate surface facilitates cell adhesion.

The small-molecule inhibitor blocks specific protein kinases, preventing cellular apoptosis.

Remove the spent medium and add fresh differentiation medium.

The medium contains a compound that promotes neural progenitor differentiation into motor neuron progenitors.

Additionally, glial committed progenitors emerge.

Next, replace with differentiation medium supplemented with a DNA synthesis inhibitor.

These inhibitors selectively induce DNA damage and death in glial progenitors, while motor neuron progenitors differentiate into mature neurons.

Wash the cells with buffer and add the differentiation medium without inhibitors to maintain the neurons.

During culturing, regularly replace the medium to replenish nutrients.

Additionally, replace the medium with medium supplemented with adhesion protein to enhance neuronal attachment, establishing a homogenous motor neuron culture.