ISOLATION OF MONOCYTES FROM BOVINE BLOOD FOR PURPOSES OF CULTURE OF DENDRITIC CELLS

Abstract

Monocytes comprise a population of mononuclear leukocytes. In this article, we compare methods for isolating monocytes and also their cultivation into dendritic cells. Bovine blood collected from the vena jugularis externa was used for the experiment. This blood was used for isolating monocytes based on density gradient of OptiPrep or Histopaque solutions. The results indicated that the more efficient method is isolation based on density gradient of the Histopaque solution and subsequent magnetic separation of the cells. The monocyte population was further cultivated for 72 h with IL-4 and GM-CSF. The cells were evaluated under a light microscope, and it was apparent that CD14-positive cells transformed to dendritic cells. The second method using the Histopaque solution and magnetic separation was more efficient for the subsequent culture of dendritic cells.