骨桥蛋白
分泌物
趋化因子
表型
细胞生物学
蛋白质组
巨噬细胞
骨髓
巨噬细胞移动抑制因子
细胞分化
四氯化碳
细胞因子
体外
生物
炎症
生物化学
免疫学
基因
作者
Rachel E. Heap,José Luis Marín‐Rubio,Julien Peltier,Tiaan Heunis,Abeer Dannoura,Adam J. Moore,Matthias Trost
标识
DOI:10.1101/2020.08.20.259515
摘要
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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