Observations on Closed Tissue Cultures of Sympathetic Ganglia of Chick Embryos in Media Buffered with N‐Tris‐(Hydroxymethyl) Methyl‐Glycine or N‐2‐Hydroxy‐Ethyl‐piperazine‐N‐2‐Ethanesulfonic Acid

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Abstract
A simple and practical closed tissue culture system for sympathetic ganglia is described. It has proved suitable in metabolic and pharmacological studies of sympathetic neurons, where it is necessary to maintain the neurons in good condition for longer periods. The effects of coverslip, Nerve Growth Factor (NGF) and various concentrations of glucose and serum on nerve fibre growth were estimated in N‐tris(hydroxymethyl)methyl‐glycine (TRICINE) or N‐2‐hydroxy‐ethylpiperazine‐N‐2‐ethanesulfonic acid (HEPES) buffered culture mediums. TRICINE and HEPES buffers were found non‐toxic to the nervous tissue and by these buffers it was possible to avoid CO2‐gassing and to use a closed environment, which facilitated daily checking of explants.Although the sympathicoblasts were not dissociated before culturing, a monolayer of the neurons with dense network of nerve fibres formed in this culture system. This pattern of growth made it possible to perform histochemical reactions (fluorescence microscopy and electron microscopical histochemical reactions) directly in the culture dishes.