Pluto Biosciences, Inc

GSE156921: Transcriptional effects of activation stimuli on M-CSF-derived macrophages

Bulk RNA sequencing

Analysis of the effect of various activation stimuli on the transcriptional signature of human M-CSF-dependent monocyte-derived macrophages.; Methods: Human Peripheral Blood Mononuclear Cells (PBMC) were isolated from buffy coats from donors over a Lymphoprep gradient according to standard procedures. Monocytes were purified from PBMC by magnetic cell sorting using anti-CD14 microbeads (>95% CD14+ cells). Monocytes from three independent donors (0.5 x 106 cells/ml, >95% CD14+ cells) were cultured in RPMI 1640 supplemented with 10% fetal bovine serum (FBS) at 37C in a humidified atmosphere with 5% CO2 and 21% O2 for 7 days in the presence of 10 ng/ml M-CSF, with cytokine addition every two days. After 7 days, macrophages were either untreated (CNT) or treated with 10 ng/ml E. coli 055:B5 lipopolysaccharide (LPS), 100 ng/ml of the TLR7 agonist CL264 (CL), BSA or 200 mM palmitate (PAL) for 0.5h, 2h, 4h or 12h. SOURCE: Miguel,A,Vega ( - CIB (CSIC)

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