Journal
JOURNAL OF IMMUNOLOGICAL METHODS
Volume 373, Issue 1-2, Pages 219-228Publisher
ELSEVIER SCIENCE BV
DOI: 10.1016/j.jim.2011.08.021
Keywords
Fetal bovine serum; Endocannabinoids; 2-Arachidonoylgylcerol; Lipids; Osteoclastogenesis; Cannabinoid receptor
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Funding
- Swiss National Science Foundation (SNSF) [3100A0-120672]
- Novartis Research Foundation [10B48]
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The major endocannabinoids (ECs) arachidonoylethanolamide (AEA) and 2-arachidonoylglycerol (2-AG) and related N-ethanolamines act as full and partial agonists at CB1, CB2, GPR55, PPAR and TRPV1 receptors to various degrees. These receptors are also expressed in immune cells like monocytes/macrophages where they regulate different cellular processes. In this study, potentially bioactive lipids in fetal bovine sera (FBS) were quantified by GC/MS. We found that several commercial FBS contain ECs and bioactive amounts of 2-AG (250-700 nM). We show that residual 2-AG from FBS can activate primary macrophages and increase migration and RANKL-stimulated osteoclastogenesis. Furthermore, 2-AG high-content sera specifically upregulated LPS-stimulated IL-6 expression in U937 cells. Polymyxin B beads may be used to selectively and efficiently remove 2-AG from sera, but not arachidonic acid and N-ethanolamines. In conclusion, 2-AG in cell culture media may significantly influence cellular experiments. CD14+ mononuclear cells which strongly express surface CB receptors may be particularly sensitive towards residual 2-AG from FBS. Therefore, the EC content in culture media should be controlled in biological experiments involving monocytes/macrophages. (C) 2011 Elsevier B.V. All rights reserved.
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