Journal of Environmental Biology

Isolation and characterization of head kidney derived

macrophages of Labeo rohita 

Abhishek Awasthi¹, Gaurav Rathore²*, P.K. Pradhan¹, S.C. Rebello¹, M.Y. Khan³ and W.S. Lakra²

¹National Bureau of Fish Genetic Resources, Lucknow-226 002, India

²Central Institute of Fisheries Education, Mumbai-400 061, India

³Baba Saheb Bhimrao Ambedkar University, Lucknow-226 025, India

*Corresponding Authors Email : grathore69@gmail.com

Publication Data

Paper received:

24 May 2013

Revised received:

05 September 2013

Accepted:

12 September 2013

Abstract

Macrophages play a significant role in non-specific defense mechanisms of all vertebrates against pathogens.? One critical element in the area of fish immunology is the unavailability of in-vitro model of immune cells. Therefore, it is essential to develop methods for harvesting and culture of macrophages for assessing innate immune functions of rohu, Labeo rohita, an important culture fish of India. Head kidney leukocytes from? were isolated by density gradient sedimentation, so as to exclude other cells. Among isolated leukocytes, only macrophages showed the unique property of sustained adherence on plastic surfaces. These cells exhibited optimum growth at 28 ?C in L-15 containing 20% FBS. Cultured head kidney macrophages (HKM) demonstrated the property of phagocytosis as evidenced by engulfment of yeast cells. Bacterial lipopolysaccharide (20?g/ml) resulted in functional activation of macrophages as seen by enhanced reactive oxygen and nitrite production; and lysosomal enzyme activity. These results show that in-vitro model of HKM cells can be used to study the role of macrophages in innate immune responses against various immunomodulators. 

Key words

Head kidney, Labeo rohita, Macrophage, Phagocytosis

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