Objective: To optimize different methods to stimulate the directional differentiation of THP-1 cells into M1, M2 macrophages and DC cells, and lay the foundation for the study of three in vitro cell models of M1, M2 and DC.
Methods: First, the differentiation of THP-1 cells was induced by stimulation with PMA and GM-CSF/M-CSF, and then different cytokines were added to induce the differentiation of THP-1 cells into M1, M2 and DC cells. Cytometry detects the expression of cell surface molecules.
RESULTS: The overall trend of the two methods to stimulate the expression of CD molecules in cells was basically the same. The expression of CD80 and CD86 on the surface of THP-1-M1 cells was significantly increased; the expression of CD163 and CD209 was highly expressed in THP-1-M2 cells; the expression of CD14 in THP-1-DC cells was significantly decreased, and CD80, CD86 and CD11c were highly expressed. After PMA stimulation, M1, M2 and DC cells all grew adherently; after GM-CSF/M-CSF stimulation, only DC cells grew adherently, while M1 and M2 cells still grew in suspension.
Conclusion: Both methods can successfully induce THP-1 cells to differentiate into different cell subtypes, but the induced cells have certain differences in morphology. The stimulation method can be selected according to the experimental needs.