Isolation of Murine Dorsal Root Ganglion Neurons

Dorsal root ganglion (DRG) neurons from mice were dissociated as previously described.^{48 (link)} Briefly, mice of either sex were anaesthetized with isoflourane and killed by cervical dislocation. The spinal cord was removed and 25–30 DRGs were extracted and washed in cold Hank’s buffered saline solution (Invitrogen). Ganglia were incubated in 900 U/ml collagenase (type XI Sigma) and 5.46 U/ml dispase (Invitrogen) for 45 min at 37°C in 5% CO₂. After that, they were gently triturated using a flame-polished glass pipette in calcium free culture medium Hank’s buffered saline solution (Invitrogen) + 1% MEM-Vit (Invitrogen) + 10% FBS (Invitrogen) + 100 mg/ml penicillin/streptomycin (Invitrogen) and the resulting suspension was centrifuged. The pellet obtained was resuspended in culture medium with the following composition: MEM (Invitrogen) + 1% MEM-Vit (Invitrogen) + 10% FBS (Invitrogen) + 100 mg/ml penicillin/streptomycin (Invitrogen). Cells were plated on round glass coverslips treated with 0.01% poly-L-lysine (Sigma) and kept at 37°C, 5% CO₂.