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Analysis of volatile organic compounds liberated and metabolised by human umbilical vein endothelial cells (HUVEC) in vitro.

Mochalski P, Theurl M, Sponring A, Unterkofler K, Kirchmair R, Amann A - Cell Biochem. Biophys. (2015)

Bottom Line: Further eight compounds (ethyl acetate, ethyl propanoate, ethyl butyrate, 3-heptanone, 2-octanone, 2-nonanone, 2-methyl-5-(methylthio)-furan and toluene) were found to be emitted by the cells under study.Possible metabolic pathways leading to the uptake and release of these compounds by HUVEC are proposed and discussed.The uptake of aldehydes by endothelial cells questions the reliability of species from this chemical class as breath or blood markers of disease processes in human organism.

View Article: PubMed Central - PubMed

Affiliation: Breath Research Institute, University of Innsbruck, Rathausplatz 4, 6850, Dornbirn, Austria, Pawel.Mochalski@uibk.ac.at.

ABSTRACT
Gas chromatography with mass spectrometric detection combined with head-space needle trap extraction as the pre-concentration technique was applied to identify and quantify volatile organic compounds released or metabolised by human umbilical vein endothelial cells. Amongst the consumed species there were eight aldehydes (2-methyl 2-propenal, 2-methyl propanal, 2-methyl butanal, 3-methyl butanal, n-hexanal, benzaldehyde, n-octanal and n-nonanal) and n-butyl acetate. Further eight compounds (ethyl acetate, ethyl propanoate, ethyl butyrate, 3-heptanone, 2-octanone, 2-nonanone, 2-methyl-5-(methylthio)-furan and toluene) were found to be emitted by the cells under study. Possible metabolic pathways leading to the uptake and release of these compounds by HUVEC are proposed and discussed. The uptake of aldehydes by endothelial cells questions the reliability of species from this chemical class as breath or blood markers of disease processes in human organism. The analysis of volatiles released or emitted by cell lines is shown to have a potential for the identification and assessment of enzymes activities and expression.

No MeSH data available.


Related in: MedlinePlus

Cultivation/measurement bottle
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Related In: Results  -  Collection


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Fig1: Cultivation/measurement bottle

Mentions: For the experiments human umbilical vein endothelial cells (HUVEC) passage 5 (P5) from pooled donors were used (PromoCell, C-12203). After building up a confluent monolayer (cell density 80–90 %) in a 75 cm2 cell culture bottle HUVEC were split 1:3 to cultivation glass bottles coated with 0.2 % gelatin solution (Sigma, G1393). The cultivation/measurement bottles had diameters of 21 cm × 5.5 cm × 11.5 cm (1,000 mL nominal volume, bottom area of approximately 240 cm2) and are shown in Fig. 1. The bottles were closed with a special Teflon plug equipped with a rubber septum enabling the insertion of the needle trap devices into the head-space of the culture and the Teflon tube being the inlet of the zero air. The inner end of the Teflon tube protruded 15–17 cm from the plug into the head-space volume, whereas the outer end was equipped with a sterile filter. The cell culture medium (EBM-2, CC-3156, supplemented with EGM-2 single quotes, CC-4147; both Clonetics) was changed every other day. After building up a confluent monolayer in the glass bottles, cells were washed three times with PBS (PAA, H15-002) and cultured in 30 mL medium. A glass bottle coated with gelatin solution (no cells) and filled with medium was used as background control. HUVEC were incubated for 24–30 h in a humidified atmosphere (37 °C and 5 % CO2) and consequently processed for the GC–MS analyses of the head-space composition. For the 24 to 30 h of cultivation, the bottles were tightly closed to boost the accumulation of species released by the cells and to block the gas exchange with the ambient air. Cell viability counts (trypan blue exclusion method) were performed immediately after the measurements. In total, 7 experiments (each involving 1 cell culture and 1 control) were performed.Fig. 1


Analysis of volatile organic compounds liberated and metabolised by human umbilical vein endothelial cells (HUVEC) in vitro.

Mochalski P, Theurl M, Sponring A, Unterkofler K, Kirchmair R, Amann A - Cell Biochem. Biophys. (2015)

Cultivation/measurement bottle
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4289529&req=5

Fig1: Cultivation/measurement bottle
Mentions: For the experiments human umbilical vein endothelial cells (HUVEC) passage 5 (P5) from pooled donors were used (PromoCell, C-12203). After building up a confluent monolayer (cell density 80–90 %) in a 75 cm2 cell culture bottle HUVEC were split 1:3 to cultivation glass bottles coated with 0.2 % gelatin solution (Sigma, G1393). The cultivation/measurement bottles had diameters of 21 cm × 5.5 cm × 11.5 cm (1,000 mL nominal volume, bottom area of approximately 240 cm2) and are shown in Fig. 1. The bottles were closed with a special Teflon plug equipped with a rubber septum enabling the insertion of the needle trap devices into the head-space of the culture and the Teflon tube being the inlet of the zero air. The inner end of the Teflon tube protruded 15–17 cm from the plug into the head-space volume, whereas the outer end was equipped with a sterile filter. The cell culture medium (EBM-2, CC-3156, supplemented with EGM-2 single quotes, CC-4147; both Clonetics) was changed every other day. After building up a confluent monolayer in the glass bottles, cells were washed three times with PBS (PAA, H15-002) and cultured in 30 mL medium. A glass bottle coated with gelatin solution (no cells) and filled with medium was used as background control. HUVEC were incubated for 24–30 h in a humidified atmosphere (37 °C and 5 % CO2) and consequently processed for the GC–MS analyses of the head-space composition. For the 24 to 30 h of cultivation, the bottles were tightly closed to boost the accumulation of species released by the cells and to block the gas exchange with the ambient air. Cell viability counts (trypan blue exclusion method) were performed immediately after the measurements. In total, 7 experiments (each involving 1 cell culture and 1 control) were performed.Fig. 1

Bottom Line: Further eight compounds (ethyl acetate, ethyl propanoate, ethyl butyrate, 3-heptanone, 2-octanone, 2-nonanone, 2-methyl-5-(methylthio)-furan and toluene) were found to be emitted by the cells under study.Possible metabolic pathways leading to the uptake and release of these compounds by HUVEC are proposed and discussed.The uptake of aldehydes by endothelial cells questions the reliability of species from this chemical class as breath or blood markers of disease processes in human organism.

View Article: PubMed Central - PubMed

Affiliation: Breath Research Institute, University of Innsbruck, Rathausplatz 4, 6850, Dornbirn, Austria, Pawel.Mochalski@uibk.ac.at.

ABSTRACT
Gas chromatography with mass spectrometric detection combined with head-space needle trap extraction as the pre-concentration technique was applied to identify and quantify volatile organic compounds released or metabolised by human umbilical vein endothelial cells. Amongst the consumed species there were eight aldehydes (2-methyl 2-propenal, 2-methyl propanal, 2-methyl butanal, 3-methyl butanal, n-hexanal, benzaldehyde, n-octanal and n-nonanal) and n-butyl acetate. Further eight compounds (ethyl acetate, ethyl propanoate, ethyl butyrate, 3-heptanone, 2-octanone, 2-nonanone, 2-methyl-5-(methylthio)-furan and toluene) were found to be emitted by the cells under study. Possible metabolic pathways leading to the uptake and release of these compounds by HUVEC are proposed and discussed. The uptake of aldehydes by endothelial cells questions the reliability of species from this chemical class as breath or blood markers of disease processes in human organism. The analysis of volatiles released or emitted by cell lines is shown to have a potential for the identification and assessment of enzymes activities and expression.

No MeSH data available.


Related in: MedlinePlus