In vitro culture requirements of sea bass (Dicentrarchus labrax L.) blood cells: Differential adhesion and phase contrast microscopic study

Abstract

The culture requirements of sea bass (Dicentrarchus labrax L.) blood leucocytes and their characterisation by phase contrast microscopy are reported. Blood leucocytes were isolated by centrifugation, maintained in primary culture and characterised. Different initial cell inocula, culture media, osmolarities and incubation temperatures were tested. Heparin and a blood dilution of 1:3 in culture medium are an indispensable condition for avoiding cell aggregates. The optimal culture conditions were incubation at 25°C after initial seeding an inoculum of 5×106cells in 2ml in a 3·5 diameter tissue culture Petri dish using RPMI-1640 culture medium. The first cell type to attach itself to the culture surface was the thrombocyte while the longest living was the monocyte/macrophage. The cultured cells show low sensitivity to osmolarity changes and a high sensitivity to incubation temperature. Non-adherent leucocytes were short-lived, and the use of a very rich culture medium did not increase cell viability.