Mouse Bone Marrow Cell Isolation and Macrophage Differentiation.

Autor:	Mendoza R; Department of Human and Molecular Genetics, Virginia Commonwealth University, Richmond, VA, USA., Banerjee I; Department of Human and Molecular Genetics, Virginia Commonwealth University, Richmond, VA, USA., Manna D; Department of Human and Molecular Genetics, Virginia Commonwealth University, Richmond, VA, USA., Reghupaty SC; C. Kenneth and Dianne Wright Center for Clinical and Translational Research, Virginia Commonwealth University, Richmond, VA, USA., Yetirajam R; Department of Human and Molecular Genetics, Virginia Commonwealth University, Richmond, VA, USA., Sarkar D; Department of Human and Molecular Genetics, Virginia Commonwealth University, Richmond, VA, USA. devanand.sarkar@vcuhealth.org.; Massey Cancer Center, Virginia Commonwealth University, Richmond, VA, USA. devanand.sarkar@vcuhealth.org.; VCU Institute of Molecular Medicine, Virginia Commonwealth University, Richmond, VA, USA. devanand.sarkar@vcuhealth.org.
Jazyk:	angličtina
Zdroj:	Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2022; Vol. 2455, pp. 85-91.
DOI:	10.1007/978-1-0716-2128-8_8
Abstrakt:	The rapid increase in the incidence of obesity contributes to a parallel increase in nonalcoholic steatohepatitis (NASH). Monocyte-derived macrophages, recruited from the bone marrow to the liver, promote NASH-related inflammation and fibrosis. In addition, adipose tissue macrophages (ATMs) release pro-inflammatory cytokines (PICs) which stimulate adipose tissue lipolysis liberating free fatty acids (FFAs) that can accumulate in the liver as triglycerides (TGs), thereby inducing steatosis. As such, bone marrow-derived macrophages (BMDMs) function as an essential tool to study the pathogenesis of NASH. BMDMs are primary bone marrow-derived cells which are differentiated into macrophages in vitro in the presence of growth factors. Macrophage colony-stimulating factor (M-CSF) is required for the proliferation and differentiation of committed myeloid progenitors into cells of the macrophage/monocyte lineage. Here, we describe a protocol for the isolation of mouse bone marrow cells and subsequent macrophage differentiation in which bone marrow cells are cultured in the presence of M-CSF, supplemented either by conditioned medium from L929 cells or in purified form. The efficiency of the differentiation is confirmed by immunofluorescent staining of macrophage surface antigen F4/80. The BMDMs serve as an excellent ex vivo model for a variety of studies, including hepatocyte-macrophage and adipocyte-macrophage cross-talk regulating NASH. (© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)
Databáze:	MEDLINE
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