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Lookup NU author(s): Rachel Heap, Dr Jose Luis Marin-RubioORCiD, Dr Julien Peltier, Dr Tiaan Heunis, Dr Abeer Dannoura, Adam Moore, Professor Matthias TrostORCiD
This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0).
© 2021 Heap et al.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.
Author(s): Heap RE, Marin-Rubio JL, Peltier J, Heunis T, Dannoura A, Moore A, Trost M
Publication type: Article
Publication status: Published
Journal: Life Science Alliance
Year: 2021
Volume: 4
Issue: 6
Print publication date: 01/06/2021
Online publication date: 14/04/2021
Acceptance date: 07/04/2021
Date deposited: 05/05/2021
ISSN (electronic): 2575-1077
Publisher: Life Science Alliance LLC
URL: https://doi.org/10.26508/lsa.202000957
DOI: 10.26508/lsa.202000957
PubMed id: 33853969
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