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Updated: Jun 5, 2026
Isolation and Culture of Bone Marrow-Derived Macrophages from Mice
Published on: June 23, 2023
Isolation and phenotypic characterization of a multinucleated tartrate-resistant acid phosphatase-positive bone
Erik Karlström1, Barbro Ek-Rylander, Mikael Wendel
1Department of Laboratory Medicine, Division of Pathology, Karolinska Institutet, Stockholm, Sweden.
Objective:
Prothrombin (PT) and osteopontin (OPN) promote adhesion of bone-derived tartrate-resistant acid phosphatase (TRAP, Acp5)-positive multinucleated cells differing in size, morphology, and resorptive activity. Here we explored phenotypic and functional differences between these cells.
Materials And Methods:
Global-wide and TRAP (Acp5) promoter messenger RNA expression, ability for phagocytosis, macrophage colony-stimulating factor-dependent migration, and in situ localization of these cells were investigated.
Results:
Gene expression of PT-adherent cells was skewed toward expression of innate immune response, phagocytosis, and scavenger receptor genes. They avidly phagocytosed Staphylococcus aureus and Dextran particles, and their migration on PT was enhanced by macrophage colony-stimulating factor. In contrast, OPN-adherent cells lacked ability to phagocytose particles and their migration on OPN was inhibited by macrophage colony-stimulating factor. They expressed typical osteoclast proteases implicated in bone matrix degradation, such as the collagenases cathepsin K; matrix metalloprotease-2; -9; -13; and -14, consistent with their high bone resorptive activity in vitro. In addition, OPN-adherent cells predominantly expressed the PU.1/MiTF/NFATc1-driven TRAP exon 1C messenger RNA, whereas PT-adherent cells preferably expressed TRAP exon 1B messenger RNA. Furthermore, CD163/cathepsin-K immunohistochemistry demonstrated that PT-adherent cells were predominantly located in the diaphyseal bone marrow compartment, whereas the OPN-adherent cells were attached to cortical and distal metaphyseal trabecular bone surfaces.
Conclusions:
This study identifies differences in expression of several osteoclast and macrophage genes, as well as functional differences between PT- and OPN-adherent cells. We conclude that the OPN-adherent cells display osteoclast characteristics, especially with regard to expression of matrix-degrading enzymes, whereas the PT-adherent cell might represent a unique TRAP-positive multinucleated bone marrow macrophage implicated in immune recognition and phagocytosis.
