Mouse resident peritoneal cells were separated into twelve fractions on Percoll gradients according to their specific density and were thoroughly characterized by Giemsa staining, some biochemical assays, immunophenotyping and phagocytic tests. Among these fractions, the macrophages were mainly represented in 7 subsets of 1.073 to 1.104 g/ml densities. The results of this study emphasize that resident peritoneal macrophages of primo-explantation can be divided into two distinct subpopulations with separate functions, related to the stage of cell maturity. In fact, our results show that one macrophage subpopulation is rich in immature cells, characterized by their peroxidative activity, the expression of F4/80 antigen, Mac-1 and Fc receptors, in correlation with their high specific density; the second subpopulation contains mature macrophages (lower percentage of peroxidase-positive cells) with lower densities and a lower level of expression of the above-mentioned molecules. Antibody-dependent and antibody-independent bacterial phagocytosis, the phagocytic index and Fc gamma RII rosetting increased together with the cell density, and were elevated in the immature cell subpopulation. T and B lymphocytes were also identified in all the macrophage subsets, but in a low proportion.