Primary culture of the hyaline haemocytes from marine decapods

Abstract

To address the dearth of techniques for medium term primary culture of shellfish blood cells, a simple method has been devised for the maintenance of crab,Liocarcinus depurator(L) andCarcinus maenas(L), hyaline haemocytes in monolayer culturein vitrofor a minimum of 14 days. This is based on L15 medium supplemented with 0·4m NaCl, 10% foetal calf serum and antibiotics. Separated hyaline cells kept in this medium remain ca. 90% viable after 2 days, >80% after 7 days and >70% after 14 days. More importantly, the cells retain defence functionality, as measured by phagocytic uptake of the marine bacterium,Psychrobacter immobilis, over the full 14 day incubation period. This method has potential value in a number of applications, particularly for fundamental studies of crustacean cellular immune processes, pathology or ecotoxicology.