Isolation, Culture, and Polarization of Murine Bone Marrow-Derived and Peritoneal Macrophages

Abstract

Macrophages are the most specialized phagocytic cells, and acquire specific phenotypes and functions in response to a variety of external triggers. Culture of bone marrow-derived or peritoneal macrophages from mice represents an exceptionally powerful technique to investigate macrophage phenotypes and functions in response to specific stimuli, resembling as much as possible the conditions observed in various pathophysiological settings. This chapter outlines protocols used to isolate and culture murine bone marrow-derived and peritoneal macrophages. Furthermore, we describe how these macrophages can be "polarized" to obtain specific macrophage subsets with special relevance to atherosclerosis.

Mesenchymal stem cells promote macrophage polarization toward M2b-like cells

“…So far, the majority of studies have used adherent cells from peritoneal exudates or spleen isolates [30,31]. Using the standard method for M isolation from the mouse peritoneum based on plastic adherence [22], these cultures contain at least 5-10% of non-M cells [25,32] [13,17,35]. This model has been used to answer two major questions: 1) how MSCs may modify the effect of an inductive environment; 2) whether the shift from one to another MΦ polarization state induced by the instantaneous microenvironment represents an irreversible or a reversible process.…”

“…BM-Ms comprised of a mature, naïve, homogenous cell population while, using the standard method for M isolation from the mouse peritoneum based on plastic adherence [22] …”

Inhibition of the lncRNA Mirt1 Attenuates Acute Myocardial Infarction by Suppressing NF-κB Activation

“…Peritoneal macrophages were harvested from C57BL/6 mice (8 - 10 weeks old) as described [20]. Aliquots of cell suspension containing 1 × 10 6 macrophages were added into the upper chamber of Transwell chambers (8 µm pore size membrane, Corning, NY, USA) in medium without serum.…”

Gelsolin Inhibits the Inflammatory Process Induced by LPS

“…After washing, the remaining cells were more than 90% macrophages. Macrophages were analyzed at this step or cultured in DMEM medium and maintained for several days [21, 22]. The Animal Committee of The Second Military University approved all mouse experiments.…”