Macrophages, the important cells of immune system, have exhibited distinct gene phenotypes with diverse functions in different microenvironments. In the present study, macrophages RAW264.7 (M0 macrophages) and lipopolysaccharide (LPS) plus interferon gamma (INF-γ)-treated M0 macrophages (M1 macrophages) were cultured in different lung cell-derived culture supernatants (CSs) as imitative tumor microenvironments. The lipids (mainly from cell membrane) of intact macrophages were in situ detected by matrix-assisted laser desorption/ionization-Fourier transform ion cyclotron resonance mass spectrometry. Approximately 300 of small molecules were observed in negative ion mode. Partial least square-discriminant analysis (PLS-DA) suggested that two types of the macrophages have different membrane lipid phenotypes. Changes in the levels of phosphatidylethanolamine PE(16:1/18:0), PE(18:1/18:0), PE(36:2), PE-Cer(d36:1), and PE(P-16:0/18:1) were closely associated with membrane phenotypes of macrophages. The heatmap also revealed that directional induction to classically activated macrophages (M1 macrophages) in vitro had greater impact on the membrane lipid phenotypes of macrophages than different lung cell-derived CSs. The results are consistent with the data obtained by biological technologies.
Keywords: Culture supernatant; Macrophage; Matrix-assisted laser desorption/ionization-mass spectrometry; Membrane lipid phenotype.
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