Study on M1 macrophage transfusion in the treatment of hepatocellular carcinoma in mice

LU Yitong; YANG Jiameng; WEI Tiaoxia; ZHAO Junlong; QIN Hongyan

doi:10.13276/j.issn.2097-1656.2024.07.005

Abstract：Objective To investigate the therapeutic effect of M1 bone marrow-derived macrophages(BMDMs)on the mouse model of orthotopic liver tumor.Methods ① Bone marrow cells isolated from wild mice were induced into BMDMs(M0 macrophages)in vitro,which were polarized into M1 macrophages under IFN-γ+LPS induction.An orthotopic mouse model of hepatocellular carcinoma was established by intrahepatic inoculation of Luciferase-Hepa1-6 cells.Seven days after tumor inoculation,PBS(control group),M0 and M1 macrophages were transfused through the tail vein,and the growth status of mouse liver tumors was detected by in vivo small-animal imaging.Twenty-one days after inoculation,liver tumor size and mass were isolated and observed to determine the effects of different types of macrophages on the progression of liver tumors in mice.Flow cytometry was used to analyze the changes of macrophage subsets in liver tumor-adjacent tissues.(2)Bone marrow cells of GFP transgenic mice were isolated and induced to differentiate into M0 and M1 macrophages,and then transfused into hepatocellular carcinoma mice via the tail vein.The number of GFP positive BMDMs in the liver was detected by flow cytometry and immunofluorescence at 1,3 and 7 d after transfusion.Results Compared with the control group,both M0 and M1 BMDMs transfusion promoted liver tumor growth(P＜0.05).After transfusion of M0 or M1 BMDMs,Kupffer cells and inflammatory macrophages increased in paracancerous tissues(P＜0.05).After 1× 106 M0 or M1 GFP+BMDMs were transfused via the tail vein,about 3x 103 cells remained in the liver after 1 d,and gradually decreased with time.Conclusion M1 macrophage transfusion does not achieve the purpose of treating liver tumors,but promotes the growth of liver tumors.