I'm looking to receive DRG neurones from a collaborator that need to be shipped overnight for culturing at our end for electrophysiology analysis. Has anyone experience with the best method to use, i.e. standard culture medium on ice, or something more spectacular? I appreciate that culture immediately after dissection is best, but that is simply not possible in this instance.
Sorry, my poor wording. shipping whole DRG for dissociation are our end.
Why don't you try doing a pilot test first to find out the best way? I'd try Hank's solution, DMEM supplemented with nerve growth factor, FBS at 20%, penicillin, and glutamine or any other cell culture medium for neurons. Then, I'd harvest some DRGs and let them overnight in these media at 4°C. On the other day, I'd dissociate them and perform ephys. Maybe none of the protocols would work or one would be better. But definitely would give a try before shipping whole DRGs!
Thanks and good idea to have a dry run as you suggest, but that is not really feasible for this collaboration - the tissue is precious and hence trying to avoid piloting. One thing though, I'd be concerned about shipping overnight in NGF because of the effects it has on neuronal gene expression (these are DRG from adults and so they do not require NGF for viability)
Ahhh I see it. But I suggested that maybe you could do it in your lab and find the best conditions for your collaborator (if possible), and of course taking in consideration mouse background etc so that everything could be appliable. Regarding NGF, you are right, I was just unaware of such effects. We use NGF for ND7 cells and to keep adult DRG neurons for XF SeaHorse analysis (MitoStress). In any case, why not trying protocols used for flow cytometry? I have one that is quite useful to keep DRG cells viable from tissue harvesting to running the cytometry.
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