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A continuous flow bioreactor system for high-throughput hyperpolarized metabolic flux analysis
Nichlas Vous Christensen1, Rikke Holm1, Juan D Sanchez2
1The MR Research Centre, Department of Clinical Medicine, Aarhus University, Aarhus, Denmark.
NMR in Biomedicine
|January 26, 2024
Summary
This study introduces a novel bioreactor system for simultaneous, ex vivo metabolic NMR spectroscopy on multiple biopsy samples using hyperpolarized carbon-13. The setup enables precise monitoring of cellular metabolism and therapeutic responses, with potential for high-throughput research.
Area of Science:
- Biomedical Engineering
- Metabolic Imaging
- Nuclear Magnetic Resonance Spectroscopy
Background:
- Hyperpolarized carbon-13 (13C) compounds are vital for real-time metabolic monitoring in medical imaging and spectroscopy.
- While radiological biomarkers are gaining traction, biopsies remain the gold standard for many diagnoses.
- Bioreactors offer a controlled environment for ex vivo biopsy analysis, crucial for monitoring therapeutic responses.
Purpose of the Study:
- To present a proof-of-concept bioreactor and microcoil array for simultaneous ex vivo metabolic NMR spectroscopy of multiple biopsy samples.
- To demonstrate the utility of this setup for hyperpolarized 13C experiments and metabolic rate constant extraction.
- To assess the preservation capabilities of the bioreactor for cellular and tissue samples.
Main Methods:
- Development of a novel bioreactor system with a microcoil receive array for simultaneous analysis of up to three biopsy samples.
- Utilization of hyperpolarized [1-13C]pyruvate for metabolic NMR spectroscopy experiments on ML-1 leukemic cells.
- Kinetic analysis of pyruvate-to-lactate conversion rates and assessment of sample preservation over time.
Main Results:
- The system successfully performed multisample carbon-13 hyperpolarization experiments with high reproducibility (CV ).
- The setup demonstrated statistical power to differentiate 30%-40% changes in lactate production, indicating treatment effects.
- ML-1 leukemic cells were preserved for at least 6 hours, and rat brain tissue slices for at least 1 hour in the bioreactor.
Conclusions:
- The developed bioreactor system provides a robust and user-friendly platform for high-throughput ex vivo metabolic studies using hyperpolarized 13C NMR spectroscopy.
- This technology facilitates precise monitoring of cellular metabolism and therapeutic efficacy in various tissue types.
- The setup lays the groundwork for advanced research in personalized medicine and drug development.
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