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Article

Determination of Intra- and Extracellular Metabolic Adaptations of 3D Cell Cultures upon Challenges in Real-Time by NMR

by 1,2,3, 1,2,3, 1,2,3, 1,2, 2,4,† and 1,5,*,†
1
Departments of Biomedical Research and Neuroradiology, University of Bern, Hochschulstrasse 6, 3012 Bern, Switzerland
2
Department of Clinical Chemistry, University Hospital Bern, Freiburgstrasse, 3010 Bern, Switzerland
3
Graduate School for Cellular and Biomedical Sciences, University of Bern, Mittelstrasse 43, 3012 Bern, Switzerland
4
Department of Pediatric Endocrinology, Diabetology and Metabolism, University Children’s Hospital of Bern, Freiburgstrasse, 3010 Bern, Switzerland
5
Translational Imaging Center (TIC), Swiss Institute for Translational and Entrepreneurial Medicine, 3010 Bern, Switzerland
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Academic Editor: Sun-Woong Kang
Int. J. Mol. Sci. 2022, 23(12), 6555; https://doi.org/10.3390/ijms23126555
Received: 6 April 2022 / Revised: 8 June 2022 / Accepted: 10 June 2022 / Published: 12 June 2022
(This article belongs to the Special Issue Development of Cell Culture Technology: Molecular Aspects and Beyond)
NMR flow devices provide longitudinal real-time quantitative metabolome characterisation of living cells. However, discrimination of intra- and extracellular contributions to the spectra represents a major challenge in metabolomic NMR studies. The present NMR study demonstrates the possibility to quantitatively measure both metabolic intracellular fingerprints and extracellular footprints on human control fibroblasts by using a commercially available flow tube system with a standard 5 mm NMR probe. We performed a comprehensive 3D cell culture system characterisation. Diffusion NMR was employed for intra- and extracellular metabolites separation. In addition, complementary extracellular footprints were determined. The implemented perfused NMR bioreactor system allowed the determination of 35 metabolites and intra- and extracellular separation of 19 metabolites based on diffusion rate differences. We show the reliability and sensitivity of NMR diffusion measurements to detect metabolite concentration changes in both intra- and extracellular compartments during perfusion with different selective culture media, and upon complex I inhibition with rotenone. We also demonstrate the sensitivity of extracellular footprints to determine metabolic variations at different flow rates. The current method is of potential use for the metabolomic characterisation of defect fibroblasts and for improving physiological comprehension. View Full-Text
Keywords: NMR; bioreactor; fibroblasts; metabolomic; diffusion; intracellular fingerprints; extracellular footprints NMR; bioreactor; fibroblasts; metabolomic; diffusion; intracellular fingerprints; extracellular footprints
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MDPI and ACS Style

Urzì, C.; Hertig, D.; Meyer, C.; Maddah, S.; Nuoffer, J.-M.; Vermathen, P. Determination of Intra- and Extracellular Metabolic Adaptations of 3D Cell Cultures upon Challenges in Real-Time by NMR. Int. J. Mol. Sci. 2022, 23, 6555. https://doi.org/10.3390/ijms23126555

AMA Style

Urzì C, Hertig D, Meyer C, Maddah S, Nuoffer J-M, Vermathen P. Determination of Intra- and Extracellular Metabolic Adaptations of 3D Cell Cultures upon Challenges in Real-Time by NMR. International Journal of Molecular Sciences. 2022; 23(12):6555. https://doi.org/10.3390/ijms23126555

Chicago/Turabian Style

Urzì, Christian, Damian Hertig, Christoph Meyer, Sally Maddah, Jean-Marc Nuoffer, and Peter Vermathen. 2022. "Determination of Intra- and Extracellular Metabolic Adaptations of 3D Cell Cultures upon Challenges in Real-Time by NMR" International Journal of Molecular Sciences 23, no. 12: 6555. https://doi.org/10.3390/ijms23126555

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