Development of leukocyte cell lines from the channel catfish (Ictalurus punctatus)

Abstract

Techniques are described for the generation of channel catfish long term leukocyte cell lines. These techniques include the isolation of peripheral blood leukocytes, purification of B cells by anti-immunoglobulin panning, mitogen stimulation, and in vitro maintenance and cloning of leukocyte cultures. Once stimulated in vitro, channel catfish leukocytes proliferate continuously without the need for exogenous growth factors or feeder cells. The various leukocyte cultures generated are heterogeneous and contain mixtures of monocyte/macrophage-like, T-like, or B cells. Clonal cell lines can be obtained from these mixed cultures by limiting dilution cloning in the presence of conditioned medium. A critical component of the culture medium is the use of channel catfish serum which is required for supporting and maintaining in vitro leukocyte proliferation. © 1994 Kluwer Academic Publishers.