Adult mouse dorsal root ganglia neurons in cell culture

Abstract

A method has been developed for the long-term culture of dissociated adult mouse dorsal root ganglia (DRG). Of critical importance to the success of this technique was a three-hour incubation in collagenase which softened the DRG and permitted gentle dissociation. The morphological and electrophysiological features of the dissociated adult DRG were similar to those observed in previous studies of immature (i.e., embryonic and newborn) DRG in culture and also to those of adult DRG in situ. With regard to electrophysiological work, the adult DRG neurons are superior to embryonic and newborn neurons because of their larger size and greatly increased survival in culture (no degeneration for first six days, and thereafter a relatively slow decrease). The adult neurons regenerated nerve fibers to an extent comparable to that of immature neurons. Therefore, the adult DRG cultures might be useful to study factors influencing regeneration in the adult mammalian nervous system. The adult cultures might also be useful to investigated factors influencing the aging process.

Sustained GSK3 activity markedly facilitates nerve regeneration

“…DRG neurons were isolated from wt and homo and heterozygous GSK3a S21A /GSK3b S9A as described previously 49 . DRG (T8-L6) were harvested, incubated in 0.25% trypsin/EDTA (GE Healthcare, Chalfont St Giles, UK) and 0.3% collagenase type IA (Sigma, St Louis, MO, USA) in DMEM (Life Technologies, Carlsbad, CA, USA) at 37°C and 5% CO 2 for 45 min and mechanically dissociated.…”

Chemical Interactions between Fibrosarcoma Cancer Cells and Sensory Neurons Contribute to Cancer Pain

“…Control cultures were prepared from L3-L6 DRGs of naive mice. Cultures of DRGs were prepared as described previously with slight modifications (Scott, 1977). After enzymatic and mechanical dissociation, the final cell suspension was plated at a density of 10,000 cells/25 mm 2 on laminin-coated glass coverslips (Fisher Scientific, Pittsburgh, PA) and maintained in Ham's F-12/DMEM supplemented with L-glutamine (2 mM), glucose (40 mM), penicillin (100 U/ml), streptomycin (100 g/ml), 5% horse serum, and DNAase I (0.15 mg/ml; Sigma, St. Louis, MO).…”

LRP1 controls biosynthetic and endocytic trafficking of neuronal prion protein

“…C on the cell surface Sensory neurons are in this study used because in vivo they uniquely have axons and no dendrites; thus, lacking any synaptic input, they are not denervated upon being dissociated and so survive, remain differentiated and regenerate their axons when introduced into culture from adult tissue (Scott, 1977). In dissociated culture, they extend axons over laminin-coated substrates and flattened satellite cells, while their large cell bodies float, usually without contacting other cells, into the medium above.…”