Vieira, M., Christensen, B. L., Wheeler, B. C., Feng, A. S., & Kollmar, R. (2007). Survival and stimulation of neurite outgrowth in a serum-free culture of spiral ganglion neurons from adult mice. Hearing research, 230(1-2), 17-23.
doi: 10.1016/j.heares.2007.03.005

In this study, a reliable protocol for the serum-free dissociation and culture of spiral ganglion neurons from adult mice and an important animal model for patients with post-lingual hearing loss is developed.  Pilot experiments indicated that the viability of spiral ganglion cells in vitro depended critically on the use of Hibernate medium with B27 supplement. With an optimized protocol, we obtained 2 x 10(3) neurons immediately after dissociation, or about one-fifth of those present in the intact spiral ganglion. 

The protocol for the completely serum-free dissociation and culture of spiral ganglion neurons from adult mice in Hibernate and Neurobasal media with B27 supplement is specified in the Materials and Methods, under the headings; 2.1. Dissociating spiral ganglion neurons and 2.2. Culturing dissociated spiral ganglion neurons. The formulation of serum-free Neurobasal medium can be found online: https://www.thermofisher.com/nl/en/home/technical-resources/media-formulation.251.html. The formulation of Hibernate medium is proprietary.