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Bone Marrow-Derived Macrophages (BMDM)

CH Chengmei Huang
RO Ruizhang Ou
XC Xiaoning Chen
YZ Yaxin Zhang
JL Jiexi Li
YL Yihao Liang
XZ Xiaohui Zhu
LL Lei Liu
ML Mingzhou Li
DL Dagui Lin
JQ Junfeng Qiu
GL Guanglong Liu
LZ Lingjie Zhang
YW Yuanyuan Wu
HT Huiyi Tang
YL Yanmin Liu
LL Li Liang
YD Yanqing Ding
WL Wenting Liao
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Bone marrow was isolated from femurs and tibias from C57 BL/6 mice using a 23- gauge needle. Cells were then cultured in RPMI 1640 supplemented with 10% FBS, 2.0mML-glutamine, 100 U/ml penicillin, 100 mg/ml streptomycin, non-essential amino acids, 14.2 mM b-mercaptoethanol, to which was added murine recombinant CSF1 (10 ng/ml; PeproTech, 351–02). Differentiating macrophages were cultured by transferring non-adherent cells 24 h after bone-marrow cells isolation and replenishing the medium with a fresh one every 48 h. BMDM differentiation was confirmed by flow cytometric evaluation of F4/80 expression. M0 macrophages (M0/M) were obtained by treating cultured cells with 10 ng/mL CSF1 for 5 days. Macrophages were then polarized to alternatively activated (M2/M) using 10 ng/mL IL4 (Peprotech, 214–14).

Copyright and License information: The Author(s) ©2021
Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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