Co-Culture of Schwann Cells with DRG Neurons on a Pre-Stretched Membrane

Take a silicon chamber attached to a matrix-coated, stretched silicon membrane.

Add dorsal root ganglia or DRG cell suspension and incubate.

The matrix promotes cell attachment and multiplication, while the stretched surface guides axon alignment.

Add growth inhibitors that eliminate actively dividing glial cells, allowing non-dividing neurons to survive.

Remove the medium. Add a growth medium and incubate.

The neuron senses an increased membrane stiffness in the stretched direction and orients the axon growth accordingly.

Add cultured and harvested Schwann cells to the DRG culture and incubate.

The Schwann cells attach to the regenerated axon, insulating the neuron.

This enhances the survival and functionality of neurons, establishing a co-culture.

Prior to seeding the DRG neurons, remove the PLL solution from the chamber. Rinse the PDMS surface with sterile water and air-dry it. After resuspending the cells, let the suspension sit for 1 to 2 minutes to allow the debris to settle to the bottom of the tube. Next, add 1.5 milliliters of cell suspension into each stretch chamber, and incubate it at 37 degrees Celsius with 5% CO₂.

To eliminate glial cells at one day in vitro, add 10 m