Isolation and Culture of Nociceptive Neurons

Nociceptive neurons were cultured as described previously [47 (link)]. Briefly, TG and DRG were dissected out from mice or rat of 2–4 month old and incubated with collagenase XI (0.66 mg/mL) and dispase II (3 mg/mL) (Sigma-Aldrich) in an INC-mix solution (NaCl 155 mM; K₂HPO₄ 1.5 mM; HEPES 10 mM; glucose 5 mM; at pH 7,4). Enzymatic digestion was performed for 45 min at 37°C in 5% CO₂, and cells were cultured in minimum essential media (MEM) supplemented with 10% FBS, Pen/Strep 100 mg/mL, MEM-vit (Invitrogen, Carlsbad CA). Cells were plated on 12-mm poly-L-lysine-coated glass coverslips and cultured for 2 days. Animal experiments were conducted in accordance with the principles and procedures of the National Institutes of Health Guidelines for the Care and Use of Laboratory and the Bioethical Committee of Universidad Católica del Norte, Coquimbo, and the Ethics Committee of the Biology Department, Faculty of Sciences, Universidad of Chile, Santiago, Chile.