Abstract
This chapter describes the methods for freshwater fishes' tissue culture. The inorganic constituents of sera from mammals, teleosts, and even from cyclostome fishes in freshwater environments are qualitatively and quantitatively similar. On the basis of physiological requirements and environmental preferences, fishes are categorized as coldwater or warmwater species, and the temperature factor applies to in vitro culture conditions. The growth of salmonid and other coldwater fish cells extends from about 5°C to 25°C with optima being near 20°C. Cells from warmwater fishes generally do well at 25°C to 30°C but may grow at 15°C through 35°C and higher. The initial pH of culture media should be about 7.3 to 7.4 for optimal results. Many fish cell cultures will grow, but with somewhat reduced activity, at pH 7.0 to 7.2 and as high as 7.8–8.0. Ovarian and embryonal tissues consistently provide good culture material. Immature testes, kidneys, spleen, swim bladder, heart, and liver may also be used, but young animals are preferred. Caudal peduncle, the trunk posterior to the anus, will usually yield successful cultures providing that young animals are used and that they are adequately decontaminated.
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