Proliferative Response of English Sole Splenic Leukocytes to Mitogens

Abstract

Abstract The mitogenic response and mixed leukocyte reaction (MLR) were studied to determine the ability of a benthic marine fish, English sole Pleuronectes vetulus, to produce leukoproliferative (LP) responses. The mitogenic response of English sole leukocytes was assessed for a variety of mitogens. In vitro culture conditions for English sole leukocytes that would maintain cellular viability and allow leukocytes to proliferate were initially determined with an undefined medium. Factors that were varied to create an effective mitogen assay included mitogen type (pokeweed mitogen, concanavalin A, phytohemagglutinin, and lipopolysaccharide), mitogen concentrations, incubation time and temperature, medium supplementation, and number of leukocytes per culture. A leukocyte concentration of 1 × 107 leukocytes/mL provided for the greatest proliferation; cell concentrations above or below this level resulted in less proliferation. A higher degree of proliferation occurred when the leukocytes were incubated with ...