Establishing a mouse model of M1/M2-polarized macrophages intrahepatic transplantation

Objective To establish a M1/M2-polarized macrophages intrahepatic transplantation model in mice. Methods Bone marrow-derived macrophages (BMDMs) were harvested. After carboxyfluorescein (FAM)-labeled small interfering RNA (siRNA) transfection, the transfected BMDM were incubated to induce M1 and M2 polarization, respectively. The BMDM were not induced marked M0. Then the 8×105 polarized BMDM were transfused to mouse liver through portal vein injection. The mouse liver injected phosphate buffer solution (PBS) were assigned to control group. FAM positive BMDM ware examined in liver tissues under fluorescence microscopy, and the mRNA levels of polarized-macrophages signature genes were measured by real-time quantitative polymerase chain reaction (real-time PCR) assays. Results Bone marrow cells were differentiated into F4/80 positive macrophages (97.60±1.25)%. After transfection, FAM positive M1 and M2 BMDM were successfully obtained. M1 and M2 signature genes were highly expressed in M1 and M2 BMDM, respectively.24 hours and 72 hours after M1 and M2 BMDM portal vein injection, the FAM positive M0, M1 and M2 BMDM could be detected in the liver tissues of recipient mice. Moreover, M1 and M2 signature genes were significantly upregulated in M1 and M2 BMDM injected liver, respectively. Conclusion This model provides an ideal animal model to investigate the function of macrophage polarization in liver disease. Key words: Macrophage polarization; Intrahepatic transplantation; Mouse model