Isolating Sensory Neurons and Co-Culturing with Natural Killer Cells

Take a dorsal root ganglion or DRG, a cluster of sensory neurons, in a cold, serum-containing medium to protect the neurons during subsequent processing. 

Centrifuge, then remove the supernatant.

Add a phosphate buffer containing digestive enzymes and incubate with mild agitation.

The enzymes dissociate the extracellular matrix, or ECM, and DRG tissue, loosening the cells.

Add a serum-containing medium to inactivate the enzymes.

Centrifuge, then discard the supernatant.

Resuspend the DRG and mechanically dissociate the tissue using pipettes of progressively smaller sizes to obtain a cell suspension.

Layer the cells onto a density gradient of bovine serum albumin.

Centrifuge. The neurons settle to the bottom, forming a pellet.

Discard the layers containing tissue debris.

Resuspend the neurons in a culture medium.

Add the neurons to an ECM-coated culture dish and incubate.

Remove the medium and add natural killer or NK cells.

Add a medium to co-culture and study the interaction between these cells.