Characteristic comparison of mouse primary macrophages cultured in L929 cell conditioned medium.

Wei Wang; Yi Qin; Yaru Wang; Jiejie Zou; Jing Chen; Jinwu Chen; Yan Zhang; Ming Geng; Zhongdong XU; Min Dai; Lilong Pan

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Characteristic comparison of mouse primary macrophages cultured in L929 cell conditioned medium.

Author: Wei WANG ¹ ; Yi QIN ¹ ; Yaru WANG ¹ ; Jiejie ZOU ¹ ; Jing CHEN ¹ ; Jinwu CHEN ¹ ; Yan ZHANG ¹ ; Ming GENG ¹ ; Zhongdong XU ¹ ; Min DAI ² ; Lilong PAN ³
Author Information

1. School of Life Sciences, Hefei Normal University, Hefei 230601, Anhui, China.
2. School of Pharmacy, Anhui University of Chinese Medicine, Hefei 230012, Anhui, China.
3. School of Medicine, Jiangnan University, Wuxi 214122, Jiangsu, China.
Publication Type:Journal Article
Keywords: macrophages; phagocytosis; polarization; primary culture; proliferation
MeSH: Animals; Bone Marrow Cells; physiology; Cells, Cultured; Culture Media, Conditioned; Lipopolysaccharides; metabolism; Macrophages; classification; physiology; Mice; Phagocytosis
From: Chinese Journal of Biotechnology 2020;36(7):1431-1439
CountryChina
Language:Chinese
Abstract: The purpose of this study is to provide a culture for mouse bone marrow-derived macrophages (BMDM) and peritoneal macrophages (PM) and to characterize their molecular and cellular biology. The cell number and purity from the primary culture were assessed by cell counter and flow cytometry, respectively. Morphological features were evaluated by inverted microscope. Phagocytosis by macrophages was detected by the neutral red dye uptake assay. Phenotypic markers were analyzed by real-time fluorescent quantitative PCR. Our results show that the cell number was much higher from culture of BMDM than PM, while there was no significant difference regarding the percentage of F4/80+CD11b+ cells (98.30%±0.53% vs. 94.83%±1.42%; P>0.05). The proliferation rate of BMDM was significantly higher than PM in the presence of L929 cell conditioned medium, by using CCK-8 assay. However, PM appeared to adhere to the flask wall and extend earlier than BMDM. The phagocytosis capability of un-stimulated BMDM was significantly higher than PM, as well as lipopolysaccharide (LPS)-stimulated BMDM, except the BMDM stimulated by low dose LPS (0.1 μg/mL). Furthermore, Tnfα expression was significantly higher in un-stimulated BMDM than PM, while Arg1 and Ym1 mRNA expression were significantly lower than PM. The expression difference was persistent if stimulated by LPS+IFN-γ or IL-4. Our data indicate that bone marrow can get larger amounts of macrophages than peritoneal cavity. However, it should be aware that the molecular and cellular characteristics were different between these two culture systems.