TY - JOUR T1 - Proteomics characterisation of the L929 cell supernatant and its role in BMDM differentiation JF - Life Science Alliance JO - Life Sci. Alliance DO - 10.26508/lsa.202000957 VL - 4 IS - 6 SP - e202000957 AU - Rachel E Heap AU - José Luis Marín-Rubio AU - Julien Peltier AU - Tiaan Heunis AU - Abeer Dannoura AU - Adam Moore AU - Matthias Trost Y1 - 2021/06/01 UR - https://www.life-science-alliance.org/content/4/6/e202000957.abstract N2 - BMDMs are a key model system to study macrophage biology in vitro. Commonly used methods to differentiate macrophages from BM are treatment with either recombinant M-CSF or the supernatant of L929 cells, which secrete M-CSF. However, little is known about the composition of L929 cell-conditioned media (LCCM) and how it affects the BMDM phenotype. Here, we used quantitative mass spectrometry to characterise the kinetics of protein secretion from L929 cells over a 2-wk period, identifying 2,193 proteins. Whereas M-CSF is very abundant in LCCM, we identified several other immune-regulatory proteins such as macrophage migration inhibitory factor (MIF), osteopontin, and chemokines such as Ccl2 and Ccl7 at surprisingly high abundance levels. We therefore further characterised the proteomes of BMDMs after differentiation with M-CSF, M-CSF + MIF, or LCCM, respectively. Interestingly, macrophages differentiated with LCCM induced a stronger anti-inflammatory M1 phenotype that those differentiated with M-CSF. This resource will be valuable to all researchers using LCCM for the differentiation of BMDMs. ER -