Isolation of DRG Neurons and Coculture with Schwann Cell Precursors to Generate Schwann Cells
Transcription
Take the spinal cord of a rat embryo with attached dorsal root ganglions, or DRGs, containing neurons and glial cells.
Detach individual DRGs from the cord, transfer them to a tube, and centrifuge, discarding any tissue debris.
Add enzymes to dissociate DRG neurons and glial cells.
Spin down the cells and remove any debris. Add a neuronal maintenance medium and triturate to generate a single-cell suspension.
Transfer the suspension into a microplate coated with a culture substrate, promoting cell adherence.
Introduce a purification medium and incubate. The medium's inhibitory agents eliminate dividing glial cells while non-dividing neurons survive.
Take Schwann cell-like cells, or SCLCs, in a co-culture medium. SCLCs, the precursor of Schwann cells, interact with embryonic peripheral nerve neurons.
Introduce the suspension onto the DRG neurons and incubate. Interaction of the SCLCs with DRG neurons promotes their maturation into Schwann cells.
The generated Schwann cells are ready for analysis.
