In Vitro Polarization of Murine Macrophage

1. In Vitro Polarization of Murine Macrophage Macrophages are tissue-resident professional phagocytes, which play an important role in immune-regulation. Activated macrophages are routinely classified into two different types: M1-macrophages (classic activation) and M2- macrophages (alternative activation), which perform pro- and anti-inflammatory active respectively. In vitro polarization of macrophages is achieved by stimulation in the presence of appropriate activation factors. Here we describe a comprehensive protocol to obtain specific macrophage subtypes through in vitro polarization from murine bone marrow derived and peritoneal macrophages. Isolation of peritoneal macrophages. Euthanize the mouse (6 to 12-week old) by rapid cervical dislocation, and then clean the abdomen with 70 % ethanol. Make an incision in the midline of the abdomen, and then expose the intact peritoneal wall. Fill a 5-mL syringe with cold PBS (+ 3% FBS), and carefully inject into the peritoneal cavity. Gently massage the peritoneum for several times to dislodge any attached cells into the PBS solution. Slowly withdraw peritoneal fluid using the same syringe, and collet the fluid into a 50-mL tube on ice. https://www.creative-diagnostics.com/in-vitro-polarization-of-murine-macrophage.htm