• Bone marrow-derived macrophages from C57BL/6 mice were successfully isolated and cultured, with F4/80 positivity reaching 98.1% after 7 days of M-CSF stimulation.
• Lipopolysaccharide (100 ng/mL) for 6 hours effectively induced M1 polarization, characterized by increased CD86 expression and upregulation of iNOS, IL-6, MIP-1α, and MCP-1 mRNA.
• Interleukin-4 (20 ng/mL) for 24 hours effectively induced M2 polarization, characterized by increased CD206 expression and upregulation of Ym1, IL-10, and Arg-1 mRNA.
• Western blot confirmed activation of p-STAT1 in M1 and p-STAT6 in M2, validating the signaling pathways involved in polarization.
• The established protocol provides a standardized in vitro model for studying macrophage polarization mechanisms and potential therapeutic applications.