In Vitro Culture of Peripheral Blood Leukocytes of <i>Channa punctatus</i>

Abstract

Cytokines are the most studied immunological molecules which play a crucial role in immunomodulatory processes involving functional T leukocytes. Cytokines are small cell-signaling protein molecules that are secreted by numerous cells mostly by peripheral blood leucocytes. In vitro culture of leucocytes plays an important role in the study of different cytokines and its role in immune system. We developed and optimized a simple and efficient method for in vitro culture of peripheral blood leukocytes of snakehead fish (Channa punctatus), using various parameters including culture medium, different mitogens and its concentration and incubation time. In the present study, peripheral blood leukocytes (PBLs) were isolated from the blood of C. punctatus using percoll and ficoll gradient solutions and were cultured in vitro in different culture media. The PBLs were stimulated with different mitogens such as phytohaemagglutinin (PHA), concanavalin A (ConA) and phorbol myristate acetate (PMA) to optimize the growth of peripheral blood leukocytes. Growth kinetics was observed to be comparatively more in PHA induced culture media. This study established an in vitro culture model that allowed us to study the growth kinetics of peripheral blood leukocytes in different media in response to different mitogens and incubation period.