Abstract: Objective To establish a method for inducing mouse bone marrow monocytes differentiating into Kupffer cells in vitro, for the study of Kupffer cells. Methods  Monocytes were separated from mouse bone marrow and induced by transforming growth factor 1 (TGF-β1), macrophage colony-stimulating factor (M-CSF) and delta-like protein 4 (DLL4). Induced monocyte-derived Kupffer cells (iMoKCs) were identified by real-time fluorescent quantitative polymerase chain reaction(qPCR), immunofluorescence staining and flow cytometry. Immunofluorescence staining and flow cytometry were used to evaluate the phagocytic capacity of iMoKCs. Cell counting kit-8(CCK-8) was used to evaluate the proliferation capacity. Results  Bone marrow monocytes were isolated, after 24 h combined induction, iMoKCs express C-type lectin domain family 4, member f（CLEC4F）,C-type lectin domain family 1, member b（CLEC1B） and V-set and immunoglobulin domain containing 4（VSIG4）at mRNA level, and protein CLEC4F. Phagocytic function of iMoKCs were detected by flow cytometry, and nearly 80% of total iMoKCs show bioparticle acceptance. The number of iMoKCs were detected by CCK-8, the counts were 1.4, 2.0 and 2.9 times of the initial number (0 h) at 24, 48 and 72 h.  Conclusion  These cells display the immunophenotype of Kupffer cells and show the ability of phagocytosis and proliferation induction through M-CSF, TGF-β1 and DLL4, which could replace primary Kupffer cells for in vitro research.

Key words: mouse, Kupffer cells, phagocytosis, proliferation, in vitro induction, monocytesKupffer