An in vitro culture system for peripheral blood leucocytes of a sciaenid fish

As a first step toward establishing techniques to monitor the immunological health of the red drum, Sciaenops ocellatus, a cell culture system was developed which supported mitogen-induced proliferation of peripheral blood leucocytes from this Sciaenid fish. RPMI1640 supplemented with homologous and human serum achieved the most consistent and most substantial responses. Culture media made isosmotic or hypoosmotic to red drum plasma supported higher levels of leucocyte proliferation than did hyperosmotic media. For PBL isolated from animals acclimated at 25° C, optimal mitogen-induced responses were routinely observed after two days of culture at 25° C. Responses to ConA were more cold-sensitive and more heat-tolerant than the response to LPS. Data obtained from these optimization studies suggested that the in vivo osmotic and temperature sensitivities characteristic of this warm water marine teleost are reflected in the in vitro responses of its isolated leucocytes.