Purpose: To optimize different methods to stimulate the directional differentiation of THP-1 cells into M1, M2 macrophages and DC cells, and to lay the foundation for the study of three in vitro cell models of M1, M2 and DC.
Methods: First, PMA and GM-CSF/M-CSF were used to stimulate the differentiation of THP-1 cells, and then different cytokines were added to induce differentiation into M1, M2 and DC cells. Observe the changes in cell morphology and use flow cytometry Cytometry detects the expression of cell surface molecules.
Result: The overall trend of the two methods to stimulate the expression of CD molecules in cells is basically the same. The expression of CD80 and CD86 on the surface of THP-1-M1 cells significantly increased; THP-1-M2 cells highly expressed CD163 and CD209; THP-1-DC cells expressed significantly lower CD14 expression, and highly expressed CD80, CD86 and CD11c. After PMA stimulation, M1, M2 and DC cells all grew adherently; after GM-CSF/M-CSF stimulation, only part of the DC cells adhered to the wall, 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 experimental needs.