DRGs were dissected from adult TrpV1-Cre::tdTomato mice
(12–13 weeks) into Hank’s balanced salt solution (HBSS) (Life
Technologies). DRG were dissociated in 1 mg ml−1 collagenase A plus 2.4
U ml−1 dispase II (enzymes, Roche Applied Sciences) in HEPES-buffered
saline (Sigma) for 90 min at 37 °C and then triturated down to single cell level
using glass Pasteur pipettes of decreasing size. DRGs were the centrifuged over a
10% BSA gradient and plated on laminin-coated cell culture dishes (Sigma). DRGs
were cultured 24 hours in B27-supplemented neurobasal-A medium plus 50 ng/ml NGF
(Invitrogen), 2 ng/ml GDNF (Sigma), 10uM arabinocytidine (Sigma) and
penicillin/streptomycin (Life Technologies).
(12–13 weeks) into Hank’s balanced salt solution (HBSS) (Life
Technologies). DRG were dissociated in 1 mg ml−1 collagenase A plus 2.4
U ml−1 dispase II (enzymes, Roche Applied Sciences) in HEPES-buffered
saline (Sigma) for 90 min at 37 °C and then triturated down to single cell level
using glass Pasteur pipettes of decreasing size. DRGs were the centrifuged over a
10% BSA gradient and plated on laminin-coated cell culture dishes (Sigma). DRGs
were cultured 24 hours in B27-supplemented neurobasal-A medium plus 50 ng/ml NGF
(Invitrogen), 2 ng/ml GDNF (Sigma), 10uM arabinocytidine (Sigma) and
penicillin/streptomycin (Life Technologies).