An improved preservation method for human dorsal root ganglion neurons enables wider access to human molecular pain...

The use of human dorsal root ganglion (DRG) from organ donors opens the door for research into molecular biology and physiology of human nociceptors; however, there are barriers to working with this tissue including logistical difficulties and limited access. We present an approach using Hibernate A media to store whole DRGs or dissociated neurons prior to culturing and functional testing. Dissociation of DRGs following temporary storage (4-16 h) in Hibernate A media resulted in similar neuronal and immune cell yield as acutely dissociated DRGs. Neurons derived from DRGs stored in Hibernate A media prior to dissociation exhibited similar electrophysiological properties and capsaicin responses as acutely dissociated DRG neurons. Similarly, neurons from acutely dissociated DRGs stored in Hibernate A media (16-42 h) and shipped to geographically distant laboratories produced neuronal cultures displaying comparable electrophysiological properties as acutely cultured neurons. This approach overcomes insurmountable logistical burdens and increases access to freshly recovered human DRGs.