Metabolomics Analyses of Cancer Cells in Controlled Microenvironments
The tumor microenvironment is a complex and heterogeneous milieu in which cancer cells undergo metabolic reprogramming to fuel their growth. Cancer cell lines grown in vitro using traditional culture methods represent key experimental models to gain a mechanistic understanding of tumor biology. This protocol describes the use of gas chromatography–mass spectrometry (GC-MS) to assess metabolic changes in cancer cells grown under varied levels of oxygen and nutrients that may better mimic the tumor microenvironment. Intracellular metabolite changes, metabolite uptake and release, as well as stable isotope (13C) tracer analyses are done in a single experimental setup to provide an integrated understanding of metabolic adaptation. Overall, this chapter describes some essential tools and methods to perform comprehensive metabolomics analyses.
Key wordsMetabolomics Microenvironment Hypoxia Nutrients Stable isotope Methoximation Silylation GC-MS
This work was supported by a Program Project Grant from the Terry Fox Research Institute (grant number TFF-116128 to J.S.-P.) and a grant from the Canadian Institutes of Health Research (CIHR, MOP-106603 to J.S.-P.). The Rosalind and Morris Goodman Cancer Research Centre Metabolomics Core Facility is supported by the Canada Foundation of Innovation, The John R. and Clara M. Fraser Memorial Trust, the Terry Fox Foundation (TFF-116128) and McGill University. J.S.-P is a FRQS research scholar.
- 13.Sato JD, Kan M (2001) Media for culture of mammalian cells. Curr Protoc Cell Biol Chapter 1:Unit 1.2Google Scholar