Spinal Cord Extraction: A Protocol to Isolate Murine Spinal Cord for In Vitro Studies

After euthanizing the animal in a CO2 chamber, soak the fur with 70% ethanol and allow it to dry. Pin the mouse in a prone position and then make a midline incision extending craniocaudally from the posterior neck to the lumbar spine.

Next, use forceps to raise the dermal flaps bilaterally and expose the subcutaneous tissue. Palpate the mouse skull until the craniocervical junction is identified. Use scissors angled perpendicular to the animal to cut through the cervical muscles and spine at the craniocervical junction. Then, dissect the spine caudally and use scissors to perform a complete caudal transection at the fifth lumbar vertebrae.

Next, position the mouse in a supine position and, after making an incision along the midline from the neck to the abdomen, retract the skin laterally and then cut open the peritoneum.

Cranially, open the chest wall with scissors. After removing the peritoneal and retroperitoneal organs, use forceps and a surgical blade to cut the ribs, leaving approximately 5 millimeters of ribs protruding from the vertebral column. Remove the vertebral column from the rest of the body and wash twice with cold PBS.