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Proteomics characterisation of the L929 cell supernatant and its role in BMDM differentiation
Open accessPreprint — published as 10.26508/lsa.202000957 · Paper page
Abstract
Bone marrow-derived macrophages (BMDMs) are a key model system to study macrophage biology in vitro . Commonly used methods to differentiate macrophages from bone marrow 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 BMDM phenotype. Here, we used quantitative mass spectrometry to characterise the kinetics of protein secretion from L929 cells over a two-week period, identifying 2,193 proteins. While 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.
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Metadata
- Authors
- Heap, R. E., Marín-Rubio, J. L., Peltier, J., Heunis, T., Dannoura, A., Moore, A., & Trost, M.
- Affiliations
- Newcastle University (GB)
- Corresponding author
- Trost, Matthias
- Venue
- Life Science Alliance
- Publisher
- openRxiv
- Published
- 2020-08-20
- Volume
- 4
- Issue
- 6
- Pages
- e202000957-e202000957
- ISSN
- 2575-1077 (ISSN-L)
- PMC
- PMC8091624
- OpenAlex
- W3152941924
- Cited by
- 80
- References
- 1
- Citations by year
- 2026: 11 · 2025: 23 · 2024: 20 · 2023: 15 · 2022: 8 · 2021: 3
- FWCI
- 4.28
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Life Science Alliance (publishedVersion, cc-by)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Newcastle University ePrints (Newcastle Univesity) (cc-by)
- Funders
- Wellcome Trust (215542/Z/19/Z, 215542)
- Newcastle University
- Directorate for Biological Sciences
- Biotechnology and Biological Sciences Research Council (215542/Z/19/Z)
- Topics
- Life Sciences › Immunology and Microbiology › Immunology › Immune cells in cancer
- Physical Sciences › Chemistry › Spectroscopy › Advanced Proteomics Techniques and Applications
- Life Sciences › Immunology and Microbiology › Immunology › Macrophage Migration Inhibitory Factor
- Keywords
- bone marrow-derived macrophages, macrophage differentiation, macrophage colony-stimulating factor, macrophage migration inhibitory factor, osteopontin, MCP-1, CCL7, chemokines, M1 macrophage polarization, macrophage phenotype
- MeSH terms
- Animals, Cell Differentiation, Humans, Macrophage Migration-Inhibitory Factors, Macrophages, Macrophage Colony-Stimulating Factor, Culture Media, Conditioned, Cell Culture Techniques, Proteomics
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