Hyperpolarized (13)C spectroscopy and an NMR-compatible bioreactor system for the investigation of real-time cellular metabolism.
Hyperpolarized (13)C spectroscopy and an NMR-compatible bioreactor system for the investigation of real-time cellular metabolism.
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DOI：
10.1002/mrm.22225
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发表时间：
2010-02
影响因子：
3.3
通讯作者：
Macdonald, Jeffrey M.
中科院分区：
文献类型：
--
作者：
Keshari, Kayvan R.;Kurhanewicz, John;Jeffries, Rex E.;Wilson, David M.;Dewar, Brian J.;Van Criekinge, Mark;Zierhut, Matthew;Vigneron, Daniel B.;Macdonald, Jeffrey M.
本研究的目的是将联合收割机与三维NMR兼容的生物反应器与超极化13 C NMR光谱结合，以便真实的时间探测细胞代谢。在三维核磁共振兼容的流化生物反应器中培养永生化大鼠肝癌细胞JM 1。在每次注入超极化[1- 13 C]丙酮酸盐之前和之后获得31 P光谱，随后在11.7 T下获得13 C光谱。在16.4 T上，从在均匀标记的13 C-葡萄糖中生长的细胞提取物获得1H和二维1H-1H-总相关光谱，以确定13 C标记的13 C分数富集和分布。发现JM 1细胞在二维培养物和生物反应器中均具有高的有氧糖酵解速率，23小时后，来自均匀标记的13 C-葡萄糖的85%的13 C标记物以乳酸盐或丙氨酸的形式存在。在生物反应器系统中，丙酮酸通过乳酸脱氢酶和丙氨酸氨基转移酶的通量测量值分别为12.18 ± 0.49 nmol/sec/108个细胞和2.39 ± 0.30 nmol/sec/108个细胞，在相同的生物反应器中是可重复的，并且在2天的过程中没有显著差异。虽然这项初步研究涉及永生化细胞，但这种技术组合可以扩展到治疗前后原发性良性和癌性细胞和组织的实时代谢探索。
The purpose of this study was to combine a three-dimensional NMR-compatible bioreactor with hyperpolarized 13C NMR spectroscopy in order to probe cellular metabolism in real time. JM1 (immortalized rat hepatoma) cells were cultured in a three-dimensional NMR-compatible fluidized bioreactor. 31P spectra were acquired before and after each injection of hyperpolarized [1-13C] pyruvate and subsequent 13C spectroscopy at 11.7 T. 1H and two-dimensional 1H-1H-total correlation spectroscopy spectra were acquired from extracts of cells grown in uniformly labeled 13C-glucose, on a 16.4 T, to determine 13C fractional enrichment and distribution of 13C label. JM1 cells were found to have a high rate of aerobic glycolysis in both two-dimensional culture and in the bioreactor, with 85% of the 13C label from uniformly labeled 13C-glucose being present as either lactate or alanine after 23 h. Flux measurements of pyruvate through lactate dehydrogenase and alanine aminotransferase in the bioreactor system were 12.18 ± 0.49 nmols/sec/108 cells and 2.39 ± 0.30 nmols/sec/108 cells, respectively, were reproducible in the same bioreactor, and were not significantly different over the course of 2 days. Although this preliminary study involved immortalized cells, this combination of technologies can be extended to the real-time metabolic exploration of primary benign and cancerous cells and tissues prior to and after therapy.
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影响因子：
5.7
作者：
Beloueche-Babari, M;Jackson, LE;Ronen, SM
通讯作者：
Ronen, SM
影响因子：
3.8
作者：
Droste, P.;Weitzel, M.;Wiechert, W.
通讯作者：
Wiechert, W.
DOI：
10.1111/j.1748-1716.1974.tb05719.x
发表时间：
1974-01-01
期刊：
ACTA PHYSIOLOGICA SCANDINAVICA
影响因子：
--
作者：
KARLSSON, J;HULTEN, B;SJODIN, B
通讯作者：
SJODIN, B
影响因子：
2.9
作者：
KURHANEWICZ, J;DAHIYA, R;NARAYAN, P
通讯作者：
NARAYAN, P
影响因子：
3.3
作者：
Chen, Albert P.;Albers, Mark J.;Vigneron, Daniel B.
通讯作者：
Vigneron, Daniel B.