Abstract
Macrophages play an essential role in diverse biological processes, from the immune response to inflammatory and neurodegenerative disorders, to various cancers. A macrophage subpopulation, known as tumor-associated macrophages (TAMs), has been shown to promote tumorigenesis, metastasis, and immune escape of cancer cells. Some of the pro-tumorigenic effects of TAMs are mediated via the secretion of nano-vesicles (exosomes) from macrophages to neighboring cells. In this chapter, we describe peritoneal macrophage isolation methods, polarization of TAMs, and purification and characterization of macrophage-derived exosomes.
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References
Wynn TA, Chawla A, Pollard JW (2013) Macrophage biology in development, homeostasis and disease. Nature 496:445–455
Sica A, Mantovani A (2012) Macrophage plasticity and polarization: in vivo veritas. J Clin Invest 122:787–795
Qian B-Z, Pollard JW (2010) Macrophage diversity enhances tumor progression and metastasis. Cell 141:39–51
Binenbaum Y et al (2018) Transfer of miRNA in macrophage-derived exosomes induces drug resistance in pancreatic adenocarcinoma. Cancer Res 78:5287–5299
Milman N, Ginini L, Gil Z (2019) Exosomes and their role in tumorigenesis and anticancer drug resistance. Drug Resist Updat 45:1–12
Zhang X, Goncalves R, Mosser DM (2008) The isolation and characterization of murine macrophages. Curr Protoc Immunol Chapter 14:Unit 14.1.doi: https://doi.org/10.1002/0471142735.im1401s83
Shelke GV, Lasser C, Gho YS, Lotvall J (2014) Importance of exosome depletion protocols to eliminate functional and RNA-containing extracellular vesicles from fetal bovine serum. J Extracell Vesicles 3. https://doi.org/10.3402/jev.v3.24783
Murray PJ et al (2014) Macrophage activation and polarization: nomenclature and experimental guidelines. Immunity 41:14–20
Nolan T, Hands RE, Bustin SA (2006) Quantification of mRNA using real-time RT-PCR. Nat Protoc 1:1559–1582
Thery C, Amigorena S, Raposo G, Clayton A (2006) Isolation and characterization of exosomes from cell culture supernatants and biological fluids. Curr Protoc Cell Biol Chapter 3:Unit 3.22
Li J, Liu K, Liu Y (2013) Exosomes mediate the cell-to-cell transmission of IFN-α-induced antiviral activity. Nat Immunol. 14(8):793–803. https://doi.org/10.1038/ni.2647
Lannigan J, Erdbruegger U (2017) Imaging flow cytometry for the characterization of extracellular vesicles. Methods. 112:55–67. https://doi.org/10.1016/j.ymeth.2016.09.018
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Fridman, E., Ginini, L., Gil, Z., Milman, N. (2020). The Purification and Characterization of Exosomes from Macrophages. In: Mishra, S. (eds) Immunometabolism. Methods in Molecular Biology, vol 2184. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0802-9_6
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DOI: https://doi.org/10.1007/978-1-0716-0802-9_6
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-0716-0802-9
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