Murine bone marrow-derived macrophages.

Abstract

AbstractThe molecular phagocytic lineage comprises, in order of increasing maturity, the committed macrophage precursor cell, the monoblast, promonocyte, monocyte, and the macrophage. Methods for the preparation and culture of bone marrow-derived macrophages, developed by Stanley and colleagues (1,2; Tushinski, R. J., personal communication), provide large numbers of mononuclear phagocytes that are capable of extensive cell proliferation. Since their proliferation can be stimulated by colony-stimulating factor-1 (CSF-1), granulocyte macrophage colony stimulating factor (GM-CSF), or interleukin-3 (IL-3), they represent an important primary cell source for studies of the actions and interactions of these three growth factors. The principles underlying the method are: 1. To generate and expand primitive mononuclear phagocyte precursor cells by culturing bone marrow cells in a combination of partially purified CSF-1 and IL-3 for a period of 3 d; 2. To remove contaminating red cells, fibroblasts, and mature macrophages and disrupt aggregates of proliferating cells by proteolytic digestion of the nonadherent cells at d 1 and 3 of culture; and 3. To obtain a population of mononuclear phagocytes that is relatively homogeneous with respect to their state of differentiation by recovering only those cells (i.e., monoblasts, promonocytes) that acquire the capacity to adhere to tissue-culture plastic during d 4–5 of culture. KeywordsGranulocyte Macrophage Colony Stimulate FactorMononuclear PhagocyteNonadherent CellProteolytic DigestionMature MacrophageThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.