The effect of insulin-like growth factor 1 and insulin on neutrophil polarization

We analyzed bone marrow derived neutrophils after they were left untreated, or treatment with LPS/IFNgamma (pro-inflammatory), IL4 (anti-inflammatory), insulin-like growth factor 1 and insulin to detect transcriptional changes after the different treatments. Neutrophils were isolated from bone marrow from femurs and tibias of C57Bl/6J mice (n=4) by Percoll gradient centrifugation, dissolved in medium (DMEM with 3% FCS, 1mM HEPES and 1% PenStrep) and plated in a 6-well plate 5-6 x 10*6 cells per well. Cells were left untreated or treated with 20 ng/mL IL-4, 10 ng/mL LPS and 2 ng/mL IFNγ, 10 ng/mL IGF1 or 10 or 100 ng/mL insulin for 4 hours at 37°C.