Serum-free culture of chicken embryonic sensory ganglia in a balanced salt solution with nerve growth factor for periods up to two weeks.

ABSTRACT

Culture of embryonic chicken dorsal root ganglia for periods exceeding a week generally require serum supplementation and a trophic factor such as nerve growth factor. In this communication we describe the culture of chicken E-9 dorsal root ganglia for periods up to 2 weeks in a system that is composed only of a simple basic salts solution and 10 ng/ml (3.8 X 10(-10)M) beta nerve growth factor. Commercially available tissue culture dishes are used which have a hydrophilic, gas-diffusable membrane on the bottom to which the ganglia directly attach, eliminating the need for added substratum. Sparse fiber outgrowth appears after 24 hours and growth of the fibers continues for 120 hours of incubation. At this time, the fibers are extremely dense and reach approximately 3.5-4.0 diameters from the body of the ganglion. Continued survival of these fibers appears to depend on the concentration of nonneuronal cells present in the dish. No fiber outgrowth occurs at any time in the absence of beta nerve growth factor. The simplicity and purity of this culture system makes it an attractive tool in the study of primary cell-cell interactions, the production of trophic factors by nonneuronal cells, and biochemical and physiological analyses of growing neurons.