Phase contrast and electron microscopical observations of adult mouse dorsal root ganglion cells maintained in primary culture
Smith, R.A.; McInnes, I.B.
Journal of Anatomy 145: 1-12
1986
ISSN/ISBN: 0021-8782 PMID: 3323151 Document Number: 275844
Cultures of neurons and non-neuronal cells were prepared from adult mouse dorsal root ganglia and maintained for periods of up to six weeks upon uncoated plastic substrates. Modifications were made to both existing isolation procedures and culture medium to ensure high cell survival. Thoracic and lumbar ganglia, excised from 3-6 month old CBA mice, were incubated overnight in 0.125% collagenase solution, thus reducing the need for excessive subsequent mechanical disruption. The isolated cells and remaining ganglion fragments were further maintained in Dulbecco's medium supplemented with 15% foetal calf serum, 10g/l glucose and 0.5% gentamycin at 37.degree. C in 5% CO2/95% air. Cell morphology was observed using phase contrast microscopy for living cells and standard light microscopy for Palmgren silver stained preparations. Scanning and transmission electron microscopy revealed the fine structure of the cells and showed the close relationship of satellite cells with the surviving dark neurons as culture establishment progressed. Satellite cells were associated with both the cell bodies and with regenerated neuritic processes. The use of cultures for future cytotoxicity testing is assessed.