Isolation and characterization of glomerular macrophages in experimental glomerulonephritis

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Abstract
Adherent cells emigrating from glomeruli of rabbits developing anti-glomerular basement membrane antibody induced glomerulonephritis were isolated and characterized as macrophages. Glomeruli were isolated using a sterile graded sieving technique and cultured in plastic tissue culture flasks. After varying culture times, emigrating adherent cells were harvested by 'cold shock' or trypsin-versene. These cells had the morphological and functional characteristics of macrophages. They were largely mononuclear, esterase-positive, phagocytic cells, which exhibited surface Fc receptors. A mean of 4821 104 macrophages could be isolated from 2 104 glomeruli after 1 h in tissue culture. Greater numbers of macrophages could be isolated with further time in culture. After 72 h however, intrinsic glomerular cell contamination occurred. The majority of the cells were viable by fluorescein diacetate hydrolysis, and Trypan Blue exclusion. Further functional studies of these cells may provide some new insights into the cellular basis of macrophage-induced glomerular injury in experimental glomerulonephritis