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Acetaldehyde and hexanaldehyde from cultured white cells.

Shin HW, Umber BJ, Meinardi S, Leu SY, Zaldivar F, Blake DR, Cooper DM - J Transl Med (2009)

Bottom Line: The mean +/- SD concentration of acetaldehyde in the headspace above the cultured cells following 4-, 24- and 48-h incubation was 157 +/- 13 ppbv, 490 +/- 99 ppbv, 698 +/- 87 ppbv.For hexanaldehyde these values were 1 +/- 0.3 ppbv, 8 +/- 2 ppbv, and 11 +/- 2 ppbv.In addition, our experimental system permitted us to identify confounding trace gas contaminants such as styrene.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biomedical Engineering, University of California, Irvine, Irvine, CA 92697, USA. hyewons@uci.edu

ABSTRACT

Background: Noninvasive detection of innate immune function such as the accumulation of neutrophils remains a challenge in many areas of clinical medicine. We hypothesized that granulocytes could generate volatile organic compounds.

Methods: To begin to test this, we developed a bioreactor and analytical GC-MS system to accurately identify and quantify gases in trace concentrations (parts per billion) emitted solely from cell/media culture. A human promyelocytic leukemia cell line, HL60, frequently used to assess neutrophil function, was grown in serum-free medium.

Results: HL60 cells released acetaldehyde and hexanaldehyde in a time-dependent manner. The mean +/- SD concentration of acetaldehyde in the headspace above the cultured cells following 4-, 24- and 48-h incubation was 157 +/- 13 ppbv, 490 +/- 99 ppbv, 698 +/- 87 ppbv. For hexanaldehyde these values were 1 +/- 0.3 ppbv, 8 +/- 2 ppbv, and 11 +/- 2 ppbv. In addition, our experimental system permitted us to identify confounding trace gas contaminants such as styrene.

Conclusion: This study demonstrates that human immune cells known to mimic the function of innate immune cells, like neutrophils, produce volatile gases that can be measured in vitro in trace amounts.

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Related in: MedlinePlus

Chromatogram of acetaldehyde from the bioreactor headspace of cells from 4-, 24- and 48-h incubations and ambient lab air. For clarity, media chromatograms are not shown (see Fig 2 for associated media responses and standard deviations). Acetaldehyde was not present in appreciable concentrations in any of the identified sources of contamination such as Teflon vials, plastic culture ware and room air samples.
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Figure 3: Chromatogram of acetaldehyde from the bioreactor headspace of cells from 4-, 24- and 48-h incubations and ambient lab air. For clarity, media chromatograms are not shown (see Fig 2 for associated media responses and standard deviations). Acetaldehyde was not present in appreciable concentrations in any of the identified sources of contamination such as Teflon vials, plastic culture ware and room air samples.

Mentions: The most prominent and reproducible signal from HL60 culture was acetaldehyde. Figure 2(A) illustrates a significantly increased emission (p < 0.0001) of acetaldehyde at 24-h and 48-h compared to 4-h from HL60 cells (4-h 157 ± 13 ppbv, 24-h 490 ± 99 ppbv and 48-h 698 ± 87 ppbv), but not from the control such as media (4-h 100 ± 9 ppbv, 24-h 170 ± 8 ppbv and 48-h 202 ± 1 ppbv). The elevated acetaldehyde observed for the HL60 was significantly higher when compared with media (p < 0.0001). Figure 2(B) illustrates the insignificant levels of acetaldehyde in all other controls (i.e., room samples, empty Teflon vial, and empty culture flasks. Figure 3 is a representative chromatogram illustrating the time-dependent increase of acetaldehyde concentration in the headspace above the HL60 cells. The asymmetry of the acetaldehyde peak is a result of the oxygenate's interaction with the column, canister and manifold. Its slower desorption from the active sites of these surfaces leads to the observed tailing [30]. The asymmetry is not observed in hexanaldehyde as its behavior is dominated by its longer hydrophobic carbon tail.


Acetaldehyde and hexanaldehyde from cultured white cells.

Shin HW, Umber BJ, Meinardi S, Leu SY, Zaldivar F, Blake DR, Cooper DM - J Transl Med (2009)

Chromatogram of acetaldehyde from the bioreactor headspace of cells from 4-, 24- and 48-h incubations and ambient lab air. For clarity, media chromatograms are not shown (see Fig 2 for associated media responses and standard deviations). Acetaldehyde was not present in appreciable concentrations in any of the identified sources of contamination such as Teflon vials, plastic culture ware and room air samples.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2683805&req=5

Figure 3: Chromatogram of acetaldehyde from the bioreactor headspace of cells from 4-, 24- and 48-h incubations and ambient lab air. For clarity, media chromatograms are not shown (see Fig 2 for associated media responses and standard deviations). Acetaldehyde was not present in appreciable concentrations in any of the identified sources of contamination such as Teflon vials, plastic culture ware and room air samples.
Mentions: The most prominent and reproducible signal from HL60 culture was acetaldehyde. Figure 2(A) illustrates a significantly increased emission (p < 0.0001) of acetaldehyde at 24-h and 48-h compared to 4-h from HL60 cells (4-h 157 ± 13 ppbv, 24-h 490 ± 99 ppbv and 48-h 698 ± 87 ppbv), but not from the control such as media (4-h 100 ± 9 ppbv, 24-h 170 ± 8 ppbv and 48-h 202 ± 1 ppbv). The elevated acetaldehyde observed for the HL60 was significantly higher when compared with media (p < 0.0001). Figure 2(B) illustrates the insignificant levels of acetaldehyde in all other controls (i.e., room samples, empty Teflon vial, and empty culture flasks. Figure 3 is a representative chromatogram illustrating the time-dependent increase of acetaldehyde concentration in the headspace above the HL60 cells. The asymmetry of the acetaldehyde peak is a result of the oxygenate's interaction with the column, canister and manifold. Its slower desorption from the active sites of these surfaces leads to the observed tailing [30]. The asymmetry is not observed in hexanaldehyde as its behavior is dominated by its longer hydrophobic carbon tail.

Bottom Line: The mean +/- SD concentration of acetaldehyde in the headspace above the cultured cells following 4-, 24- and 48-h incubation was 157 +/- 13 ppbv, 490 +/- 99 ppbv, 698 +/- 87 ppbv.For hexanaldehyde these values were 1 +/- 0.3 ppbv, 8 +/- 2 ppbv, and 11 +/- 2 ppbv.In addition, our experimental system permitted us to identify confounding trace gas contaminants such as styrene.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Biomedical Engineering, University of California, Irvine, Irvine, CA 92697, USA. hyewons@uci.edu

ABSTRACT

Background: Noninvasive detection of innate immune function such as the accumulation of neutrophils remains a challenge in many areas of clinical medicine. We hypothesized that granulocytes could generate volatile organic compounds.

Methods: To begin to test this, we developed a bioreactor and analytical GC-MS system to accurately identify and quantify gases in trace concentrations (parts per billion) emitted solely from cell/media culture. A human promyelocytic leukemia cell line, HL60, frequently used to assess neutrophil function, was grown in serum-free medium.

Results: HL60 cells released acetaldehyde and hexanaldehyde in a time-dependent manner. The mean +/- SD concentration of acetaldehyde in the headspace above the cultured cells following 4-, 24- and 48-h incubation was 157 +/- 13 ppbv, 490 +/- 99 ppbv, 698 +/- 87 ppbv. For hexanaldehyde these values were 1 +/- 0.3 ppbv, 8 +/- 2 ppbv, and 11 +/- 2 ppbv. In addition, our experimental system permitted us to identify confounding trace gas contaminants such as styrene.

Conclusion: This study demonstrates that human immune cells known to mimic the function of innate immune cells, like neutrophils, produce volatile gases that can be measured in vitro in trace amounts.

Show MeSH
Related in: MedlinePlus