Metabolomic and Lipidomic Analysis of Bone Marrow Derived Macrophages.
| Autor: | Seim GL; Morgridge Institute for Research, Madison, WI, USA.; Department of Nutritional Sciences, University of Wisconsin-Madison, Madison, WI, USA., John SV; Morgridge Institute for Research, Madison, WI, USA., Fan J; Morgridge Institute for Research, Madison, WI, USA.; Department of Nutritional Sciences, University of Wisconsin-Madison, Madison, WI, USA. |
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| Jazyk: | angličtina |
| Zdroj: | Bio-protocol [Bio Protoc] 2020 Jul 20; Vol. 10 (14), pp. e3693. Date of Electronic Publication: 2020 Jul 20 (Print Publication: 2020). |
| DOI: | 10.21769/BioProtoc.3693 |
| Abstrakt: | Macrophages are highly plastic immune cells that are capable of adopting a wide array of functional phenotypes in response to environmental stimuli. The changes in macrophage function are often supported and regulated by changes in cellular metabolism. Capturing a comprehensive picture of metabolism is vital for understanding the role of metabolic rewiring in the immune response. Here we present a method for systematically quantifying the abundance of metabolites and lipids in primary murine bone marrow derived macrophages (BMDMs). This method simultaneously extracts polar metabolites and lipids from BMDMs using a rapid two-phase extraction procedure. The polar metabolite fraction and lipid fraction are subsequently analyzed by separate liquid chromatography-mass spectrometry (LC-MS) methods for optimized coverage and quantification. This allows for a comprehensive characterization of cellular metabolism that can be used to understand the impact of a variety of environmental stimuli on macrophage metabolism and function. Competing Interests: Competing interestsThe authors declare no competing interests. (Copyright © 2020 The Authors; exclusive licensee Bio-protocol LLC.) |
| Databáze: | MEDLINE |
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