Cost-Effective Real-Time Metabolic Profiling of Cancer Cell Lines for Plate-Based Assays
Open accessAbstract
A fundamental phenotype of cancer cells is their metabolic profile, which is routinely described in terms of glycolytic and respiratory rates. Various devices and protocols have been designed to quantify glycolysis and respiration from the rates of acid production and oxygen utilization, respectively, but many of these approaches have limitations, including concerns about their cost-ineffectiveness, inadequate normalization procedures, or short probing time-frames. As a result, many methods for measuring metabolism are incompatible with cell culture conditions, particularly in the context of high-throughput applications. Here, we present a simple plate-based approach for real-time measurements of acid production and oxygen depletion under typical culture conditions that enable metabolic monitoring for extended periods of time. Using this approach, it is possible to calculate metabolic fluxes and, uniquely, describe the system at steady-state. By controlling the conditions with respect to pH buffering, O2 diffusion, medium volume, and cell numbers, our workflow can accurately describe the metabolic phenotype of cells in terms of molar fluxes. This direct measure of glycolysis and respiration is conducive for between-runs and even between-laboratory comparisons. To illustrate the utility of this approach, we characterize the phenotype of pancreatic ductal adenocarcinoma cell lines and measure their response to a switch of metabolic substrate and the presence of metabolic inhibitors. In summary, the method can deliver a robust appraisal of metabolism in cell lines, with applications in drug screening and in quantitative studies of metabolic regulation.
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Supplementation with IL-6 or hyIL-6 prior to metabolic phenotyping has no added effect in MIA PaCa-2 cells.
“supplementation with IL-6 or its synthetic fusion protein, hyper-IL-6 (hyIL-6), 24 prior to metabolic phenotyping had no added effect”
From Dynamic IL-6R/STAT3 signaling leads to heterogeneity of metabolic phenotype in pancreatic ductal adenocarcinoma cells (2024) · cites this paper as [24]
Section 3.4.3 describes metabolic measurements in MIA-PaCa-2 cells after changing the substrate or adding rotenone. The Appendix figure captions likewise describe substrate and rotenone experiments, but neither these passages nor searches for IL-6 and hyIL-6 address cytokine supplementation or whether it had no added effect. No page number is provided in our copy for these passages. (LLM, conf=0.76)
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Self-citation
- Authors
- Blaszczak, W., Tan, Z., & Swietach, P.
- Affiliations
- Department of Physiology, Anatomy & Genetics, University of Oxford, Oxford OX1 3PT, UK
- University of Oxford (GB)
- Corresponding author
- Swietach, Pawel
- Venue
- Chemosensors
- Publisher
- MDPI AG
- Published
- 2021-06-13
- Volume
- 9
- Issue
- 6
- Pages
- 139
- ISSN
- 2227-9040 (ISSN-L)
- OpenAlex
- W3165947290
- Cited by
- 10
- References
- 44
- Citations by year
- 2026: 1 · 2025: 2 · 2024: 1 · 2023: 2 · 2022: 4
- FWCI
- 0.70
- Type
- journal article
- Language
- en
- Open access
- gold
- doi.org
- Licence
- Locations
- Chemosensors (publishedVersion, cc-by)
- DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
- Spiral (Imperial College London) (submittedVersion, other-oa)
- Funders
- European Research Council (723997)
- Marie Skłodowska-Curie Innovative Training Network (813834)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cancer Research › Cancer, Hypoxia, and Metabolism
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Cancer and biochemical research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Microbial Metabolic Engineering and Bioproduction
- Keywords
- cancer metabolism, glycolysis, cellular respiration, acid production, oxygen consumption, metabolic flux, plate-based assay, metabolic inhibitors, drug screening, steady-state metabolism
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