Understanding natural isotopic variations in cultured cancer cells

IF 1.8 3区化学 Q4 BIOCHEMICAL RESEARCH METHODS

Rapid Communications in Mass Spectrometry Pub Date : 2024-08-08 DOI:10.1002/rcm.9878

Olivier L. Mantha, Marie Mahé, Karine Mahéo, Gaëlle Fromont, Maxime Guéguinou, Illa Tea, Régis Hankard, Arnaud De Luca

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引用次数: 0

Abstract

Rationale

Natural variations in the abundance of the stable isotopes of nitrogen (δ¹⁵N) and carbon (δ¹³C) offer valuable insights into metabolic fluxes. In the wake of strong interest in cancer metabolism, recent research has revealed δ¹⁵N and δ¹³C variations in cancerous compared to non-cancerous tissues and cell lines. However, our understanding of natural isotopic variations in cultured mammalian cells, particularly in relation to metabolism, remains limited. This study aims to start addressing this gap using metabolic modulations in cells cultured under controlled conditions.

Methods

Prostate cancer cells (PC3) were cultured in different conditions and their δ¹⁵N and δ¹³C were measured using isotope ratio mass spectrometry. Isotopic variations during successive cell culture passages were assessed and two widely used cell culture media (RPMI and DMEM) were compared. Metabolism was modulated through glutamine deprivation and hypoxia.

Results

Successive cell culture passages generally resulted in reproducible δ¹⁵N and δ¹³C values. The impact of culture medium composition on δ¹⁵N and δ¹³C of the cells highlights the importance of maintaining a consistent medium composition across conditions whenever possible. Glutamine deprivation and hypoxia induced a lower δ¹³C in bulk cell samples, with only the former affecting δ¹⁵N. Gaps between theory and experiments were bridged and the lessons learned throughout the process are provided.

Conclusions

Exposing cultured cancer cells to hypoxia allowed us to further investigate the relation between metabolic modulations and natural isotopic variations, while mitigating the confounding impact of changing culture medium composition. This study highlights the potential of natural δ¹³C variations for studying substrate fluxes and nutrient allocation in reproducible culture conditions. Considering cell yield and culture medium composition is pivotal to the success of this approach.

Abstract Image

查看原文本刊更多论文

了解培养癌细胞中的天然同位素变化。

理由：氮（δ15N）和碳（δ13C）稳定同位素丰度的自然变化为了解代谢通量提供了宝贵的信息。随着人们对癌症代谢的浓厚兴趣，最近的研究发现，与非癌症组织和细胞系相比，癌症组织和细胞系中的δ15N和δ13C发生了变化。然而，我们对培养的哺乳动物细胞中天然同位素变化，特别是与新陈代谢有关的同位素变化的了解仍然有限。本研究旨在利用在受控条件下培养的细胞中的代谢调节，着手解决这一空白：方法：在不同条件下培养前列腺癌细胞（PC3），并使用同位素比质谱法测量其δ15N和δ13C。评估了细胞连续培养过程中的同位素变化，并对两种广泛使用的细胞培养基（RPMI 和 DMEM）进行了比较。通过谷氨酰胺剥夺和缺氧调节新陈代谢：结果：连续培养细胞通常可获得可重复的δ15N 和 δ13C值。培养基成分对细胞δ15N和δ13C的影响凸显了在各种条件下保持培养基成分一致的重要性。谷氨酰胺剥夺和缺氧会导致大量细胞样本中的δ13C降低，只有前者会影响δ15N。我们弥补了理论与实验之间的差距，并在整个过程中吸取了经验教训：将培养的癌细胞置于缺氧环境中，使我们能够进一步研究代谢调节与天然同位素变化之间的关系，同时减轻培养基成分变化的干扰影响。这项研究凸显了天然 δ13C 变化在可重复培养条件下研究底物通量和营养分配的潜力。考虑细胞产量和培养基成分对这种方法的成功至关重要。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Rapid Communications in Mass Spectrometry 化学-分析化学

CiteScore

4.10

自引率

5.00%

发文量

219

审稿时长

2.6 months

期刊介绍： Rapid Communications in Mass Spectrometry is a journal whose aim is the rapid publication of original research results and ideas on all aspects of the science of gas-phase ions; it covers all the associated scientific disciplines. There is no formal limit on paper length ("rapid" is not synonymous with "brief"), but papers should be of a length that is commensurate with the importance and complexity of the results being reported. Contributions may be theoretical or practical in nature; they may deal with methods, techniques and applications, or with the interpretation of results; they may cover any area in science that depends directly on measurements made upon gaseous ions or that is associated with such measurements.