Proteomics characterisation of the L929 cell supernatant and its role in BMDM differentiation

• DOI: 10.26508/lsa.202000957
• Publication Date: 2021-04-14T20:46:33Z
• Authors: Rachel E Heap; José Luis Marín-R...; Julien Peltier; Tiaan Heunis; Abeer Dannoura; Adam Moore; Matthias Trost
• Keywords: Osteopontin; Proteome

BMDMs are a key model system to study macrophage biology in vitro. Commonly used methods differentiate macrophages from BM treatment with either recombinant M-CSF or the supernatant of L929 cells, which secrete M-CSF. However, little is known about composition cell-conditioned media (LCCM) and how it affects BMDM phenotype. Here, we quantitative mass spectrometry characterise kinetics protein secretion cells over 2-wk period, identifying 2,193 proteins. Whereas very abundant LCCM, identified several other immune-regulatory proteins such as migration inhibitory factor (MIF), osteopontin, chemokines Ccl2 Ccl7 at surprisingly high abundance levels. We therefore further characterised proteomes after differentiation M-CSF, + MIF, respectively. Interestingly, differentiated LCCM induced stronger anti-inflammatory M1 phenotype that those This resource will be valuable all researchers using for BMDMs.