Journal article
A novel bioreactor for combined magnetic resonance spectroscopy and optical imaging of metabolism in 3D cell cultures
Magnetic resonance in medicine, Vol.81(5), pp.3379-3391
05/2019
DOI: 10.1002/mrm.27644
PMCID: PMC6414270
PMID: 30652350
Abstract
Fluorescence lifetime imaging microscopy (FLIM) of endogenous fluorescent metabolites permits the measurement of cellular metabolism in cell, tissue and animal models. In parallel, magnetic resonance spectroscopy (MRS) of dynamic nuclear (hyper)polarized (DNP)
C-pyruvate enables measurement of metabolism at larger in vivo scales. Presented here are the design and initial application of a bioreactor that connects these 2 metabolic imaging modalities in vitro, using 3D cell cultures.
The model fitting for FLIM data analysis and the theory behind a model for the diffusion of pyruvate into a collagen gel are detailed. The device is MRI-compatible, including an optical window, a temperature control system and an injection port for the introduction of contrast agents. Three-dimensional printing, computer numerical control machining and laser cutting were used to fabricate custom parts.
Performance of the bioreactor is demonstrated for 4 T1 murine breast cancer cells under glucose deprivation. Mean nicotinamide adenine dinucleotide (NADH) fluorescence lifetimes were 10% longer and hyperpolarized
C lactate:pyruvate (Lac:Pyr) ratios were 60% lower for glucose-deprived 4 T1 cells compared to 4 T1 cells in normal medium. Looking at the individual components of the NADH fluorescent lifetime, τ
(free NADH) showed no significant change, while τ
(bound NADH) showed a significant increase, suggesting that the increase in mean lifetime was due to a change in bound NADH.
A novel bioreactor that is compatible with, and can exploit the benefits of, both FLIM and
C MRS in 3D cell cultures for studies of cell metabolism has been designed and applied.
Details
- Title: Subtitle
- A novel bioreactor for combined magnetic resonance spectroscopy and optical imaging of metabolism in 3D cell cultures
- Creators
- Benjamin L Cox - Morgridge Institute for ResearchSarah Erickson-Bhatt - University of Wisconsin–MadisonJoseph M Szulczewski - University of Wisconsin–MadisonJayne M Squirrell - University of Wisconsin–MadisonKai D Ludwig - University of Wisconsin–MadisonErin B Macdonald - University of Wisconsin–MadisonRobert Swader - Morgridge Institute for ResearchSuzanne M Ponik - University of Wisconsin–MadisonKevin W Eliceiri - University of Wisconsin–MadisonSean B Fain - University of Wisconsin–Madison
- Resource Type
- Journal article
- Publication Details
- Magnetic resonance in medicine, Vol.81(5), pp.3379-3391
- DOI
- 10.1002/mrm.27644
- PMID
- 30652350
- PMCID
- PMC6414270
- NLM abbreviation
- Magn Reson Med
- ISSN
- 0740-3194
- eISSN
- 1522-2594
- Grant note
- CA142833 / NIH HHS CA142833, CA185251 and CA179556. / NIH HHS CA185251 / NIH HHS UL1TR000427 and TL1TR000429 / NIH/NCATS R01 CA185251 / NCI NIH HHS UL1TR000427 / NIH/NCATS UL1 TR002373 / NCATS NIH HHS TL1TR000429 / NIH/NCATS R01 HL126771 / NHLBI NIH HHS T32 CA009206 / NCI NIH HHS TL1 TR000429 / NCATS NIH HHS T32CA00926 / NIH/NCI R01 CA179556 / NCI NIH HHS UL1 TR000427 / NCATS NIH HHS R01 CA142833 / NCI NIH HHS TL1 TR002375 / NCATS NIH HHS CA179556. / NIH HHS
- Language
- English
- Date published
- 05/2019
- Academic Unit
- Roy J. Carver Department of Biomedical Engineering; Radiology; Electrical and Computer Engineering; Health, Sport, and Human Physiology
- Record Identifier
- 9984275056302771
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