Variable VOCs in plastic culture flasks and their potential impact on cell volatile biomarkers

Chu, Yajing; Zhou, Jijuan; Ge, Dianlong; Lü, Yan; Zou, Xue; Xia, Lei; Huang, Chaoqun; Shen, Chengyin; Chu, Yannan

doi:10.1007/s00216-020-02756-9

Cited by 17 publications

(16 citation statements)

References 26 publications

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“…Correspondingly, after cell culturing, 2methyl-butanol, 3-methyl-butanol, and 2-methyl-propanol increased for the reduction of the corresponding aldehyde. This result is not surprising considering the potential impact of VOC in plastic culture flasks on cell metabolism (Filipiak et al, 2016;Chu et al, 2020). More interestingly, we observed after neuron culturing a significant increase in other nine VOCs: 3-OH-2-butanone (acetoin), 2,3-butanedione, propanal and 2methyl-propanal, methanol and methylacetate, propanol, 2pentanol, and acetone.…”

Section: Discussionsupporting

confidence: 65%

Unraveling the Extracellular Metabolism of Immortalized Hippocampal Neurons Under Normal Growth Conditions

et al. 2021

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Metabolomic profiling of cell lines has shown many potential applications and advantages compared to animal models and human subjects, and an accurate cellular metabolite analysis is critical to understanding both the intracellular and extracellular environments in cell culture. This study provides a fast protocol to investigate in vitro metabolites of immortalized hippocampal neurons HN9.10e with minimal perturbation of the cell system using a targeted approach. HN9.10e neurons represent a reliable model of one of the most vulnerable regions of the central nervous system. Here, the assessment of their extracellular metabolic profile was performed by studying the cell culture medium before and after cell growth under standard conditions. The targeted analysis was performed by a direct, easy, high-throughput reversed-phase liquid chromatography with diode array detector (RP-HPLC-DAD) method and by headspace solid-phase microextraction–gas chromatography–mass spectrometry (HS-SPME-GC-MS) for the study of volatile organic compounds (VOCs). The analysis of six different batches of cells has allowed to investigate the metabolic reproducibility of neuronal cells and to describe the metabolic “starting” conditions that are mandatory for a well-grounded interpretation of the results of any following cellular treatment. An accurate study of the metabolic profile of the HN9.10e cell line has never been performed before, and it could represent a quality parameter before any other targeting assay or further exploration.

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Section: Discussionsupporting

confidence: 65%

Unraveling the Extracellular Metabolism of Immortalized Hippocampal Neurons Under Normal Growth Conditions

et al. 2021

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“…This was apparent in PCA analysis which was confounded by date of sampling. Plasticware used to grow cells produces a high number of VOCs, particularly derivatives of benzene, such as styrene, toluene and ethylbenzene [39,48,49]. Glassware represents an alternative for use in headspace experiments that is inert and does not produce VOCs [49].…”

Section: Discussionmentioning

confidence: 99%

“…This may have led to an accumulation of exogenous contaminant VOCs as well as biologically relevant VOCs. In addition, the caps used in cell culture sampling are known to be a high producer of contaminant VOCs [48]. We performed active sampling outside of the incubator which meant that humidity, temperature, and CO 2 levels could not be regulated.…”

Section: Discussionmentioning

confidence: 99%

Changes in lung epithelial cell volatile metabolite profile induced by pro-fibrotic stimulation with TGF-β1

Hayton,

Ahmed,

Cunningham

et al. 2023

J. Breath Res.

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IntroductionVolatile organic compounds (VOCs) have shown promise as potential biomarkers in idiopathic pulmonary fibrosis (IPF). Measuring VOCs in the headspace of in vitro models of lung fibrosis may offer a method of determining the origin of those detected in exhaled breath. The aim of this study was to determine the VOCs associated with two lung cell lines (A549 and MRC-5 cells) and changes associated with stimulation of cells with the pro-fibrotic cytokine, transforming growth factor (TGF)-β1.MethodsA dynamic headspace sampling method was used to sample the headspace of A549 cells and MRC-5 cells. These were compared to media control samples and to each other to identify VOCs which discriminated between cell lines. Cells were then stimulated with the TGF-β1 and samples were compared between stimulated and unstimulated cells. Samples were analysed using thermal desorption-gas chromatography-mass spectrometry and supervised analysis was performed using sparse partial least squares-discriminant analysis (sPLS-DA).ResultsSupervised analysis revealed differential VOC profiles unique to each of the cell lines and from the media control samples. Significant changes in VOC profiles were induced by stimulation of cell lines with TGF-β1. In particular, several terpenoids (isopinocarveol, sativene and 3-carene) were increased in stimulated cells compared to unstimulated cells.ConclusionsVOC profiles differ between lung cell lines and alter in response to pro-fibrotic stimulation. Increased abundance of terpenoids in the headspace of stimulated cells may reflect TGF-β1 cell signalling activity and metabolic reprogramming. This may offer a potential biomarker target in exhaled breath in IPF. 

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“…Blank procedure: the temperature is maintained at 40 °C for 1 min, raised to 200 °C at a speed of 5 °C/min, and hold at 200 °C for 2 min. The MS operation conditions were the same as our previous work . The ionization energy was 70 eV, and the mass scanning range was 45–300 amu.…”

Section: Methodsmentioning

confidence: 99%

HS–SPME–GC–MS Untargeted Analysis of Normal Rat Organs Ex Vivo: Differential VOC Discrimination and Fingerprint VOC Identification

Liu

Zhou

et al. 2023

Anal. Chem.

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The investigation of volatile organic compounds (VOCs) in human metabolites has been a topic of interest as it holds the potential for the development of non-invasive technologies to screen for organ lesions in vivo. However, it remains unclear whether VOCs differ among healthy organs. Consequently, a study was conducted to analyze VOCs in ex vivo organ tissues obtained from 16 Wistar rats, comprising 12 different organs. The VOCs released from each organ tissue were detected by the headspace-solid phase microextraction-gas chromatography-mass spectrometry technique. In the untargeted analysis of 147 chromatographic peaks, the differential volatiles of rat organs were explored based on the Mann–Whitney U test and fold change (FC > 2.0) compared with other organs. It was found that there were differential VOCs in seven organs. A discussion on the possible metabolic pathways and related biomarkers of organ differential VOCs was conducted. Based on the orthogonal partial least squares discriminant analysis and receiver operating characteristic curve, we found that differential VOCs in the liver, cecum, spleen, and kidney can be used as the unique identification of the corresponding organ. In this study, differential VOCs of organs in rats were systematically reported for the first time. Profiles of VOCs produced by healthy organs can serve as a reference or baseline that may indicate the presence of disease or abnormalities in the organ’s function. Differential VOCs can be used as the fingerprint of organs, and future integration with metabolic research may contribute to the development of healthcare.

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Variable VOCs in plastic culture flasks and their potential impact on cell volatile biomarkers

Cited by 17 publications

References 26 publications

Unraveling the Extracellular Metabolism of Immortalized Hippocampal Neurons Under Normal Growth Conditions

Unraveling the Extracellular Metabolism of Immortalized Hippocampal Neurons Under Normal Growth Conditions

Changes in lung epithelial cell volatile metabolite profile induced by pro-fibrotic stimulation with TGF-β1

HS–SPME–GC–MS Untargeted Analysis of Normal Rat Organs Ex Vivo: Differential VOC Discrimination and Fingerprint VOC Identification

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