Academic Journal
Hyperpolarized 13C NMR for longitudinal in-cell metabolism using a mobile 3D cell culture system
| Title: | Hyperpolarized 13C NMR for longitudinal in-cell metabolism using a mobile 3D cell culture system |
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| Authors: | T.B.W. Mathiassen, A.E. Høgh, M. Karlsson, S. Katsikis, K. Wang, M. Pennestri, J.H. Ardenkjær-Larsen, P.R. Jensen |
| Source: | Journal of Magnetic Resonance Open, Vol 16, Iss , Pp 100131- (2023) |
| Publisher Information: | Elsevier, 2023. |
| Publication Year: | 2023 |
| Collection: | LCC:Medical physics. Medical radiology. Nuclear medicine LCC:Physics |
| Subject Terms: | Dissolution dynamic nuclear polarization, In-Cell NMR, Metabolism, Longitudinal cell studies, Nuclear magnetic resonance, Medical physics. Medical radiology. Nuclear medicine, R895-920, Physics, QC1-999 |
| More Details: | Hyperpolarization with the dissolution dynamic nuclear polarization (dDNP) technique yields > 10,000-fold signal increases for NMR-active nuclei (e.g. 13C). Hyperpolarized 13C-labeled metabolic tracer molecules thus allow real-time observations of biochemical pathways in living cellular systems without interfering background. This methodology lends itself to the direct observation of altered intracellular reaction chemistry imparted for instance by drug treatment, infections, or other diseases. A reoccurring challenge for longitudinal cell studies of mammalian cells with NMR and dDNP-NMR is maintaining cell viability in the NMR spectrometer. 3D cell culture methods are increasing in popularity because they provide a physiologically more relevant environment compared to 2D cell cultures. Based on such strategies a mobile 3D culture system was devised. The clinical drug etoposide was used to treat cancer cells (HeLa) and the resulting altered metabolism was measured using hyperpolarized [1–13C]pyruvate. We show that sustaining the cell cultivation in cell incubators and only transferring the cells to the NMR spectrometer for the few minutes required for the dDNP-NMR measurements is an attractive alternative to cell maintenance in the NMR tube. High cell viability is sustained, and experimental throughput is many doubled. |
| Document Type: | article |
| File Description: | electronic resource |
| Language: | English |
| ISSN: | 2666-4410 |
| Relation: | http://www.sciencedirect.com/science/article/pii/S2666441023000390; https://doaj.org/toc/2666-4410 |
| DOI: | 10.1016/j.jmro.2023.100131 |
| Access URL: | https://doaj.org/article/1a55ded60c564b53866afcce36f688c2 |
| Accession Number: | edsdoj.1a55ded60c564b53866afcce36f688c2 |
| Database: | Directory of Open Access Journals |
| ISSN: | 26664410 |
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| DOI: | 10.1016/j.jmro.2023.100131 |
| Published in: | Journal of Magnetic Resonance Open |
| Language: | English |