Figure 13.

DRG dissection and dissociation protocol. Lumbar DRGs (L1–L6) were removed from the mice and transferred to a petri dish where the nerves were trimmed for cleaning. Individual DRGs were excised to expose the cells. Enzymatic tissue dissociation was performed using a trypsin solution for 20 min at 37 °C. DRGs were then mechanically dissociated by pipetting using Pasteur pipettes with decreasing diameter tips and sieved through a 45-µm cell strainer. Cell suspension was centrifuged and pelleted cells were resuspended in 100 μL of the culture medium. Aliquots of the resuspended solution were plated in 24-well plates within a silicone ring and incubated overnight in a cell culture chamber at 37 °C.