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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

The 378 mL glass bioreactor designed for incubating cells in air containing low volatile organic compounds and post incubation collection of the gaseous headspace.
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Figure 1: The 378 mL glass bioreactor designed for incubating cells in air containing low volatile organic compounds and post incubation collection of the gaseous headspace.

Mentions: The Teflon vials containing the cell suspensions (40 × 106 cells/30 ml) were placed inside cylindrical glass bioreactors. The glass bioreactors were specifically designed to collect the gaseous headspace above aqueous cultures (see Figure 1) [19]. The bioreactor consisted of two glass halves joined together with an o-ring and secured by a spherical joint Thomas® pinch clamp. The bioreactor had an interior volume of 378 mL and was fitted with valves, sealed with high vacuum Chem-Vac™ stopcocks, at both ends. Once the apparatus was fully assembled it was attached to a pressurized manifold to purge the bioreactor of ambient air and replace it with air containing low levels of volatile organic compounds (VOCs) and 5% CO2. The low VOC air was prepared by doping 5% pure CO2 in to whole air collected by the Blake-Rowland lab from the rural Crooked Creek Research Station in California's White Mountains [29]. Figure 2(B) and 4(B) illustrate the low levels of selected VOCs in the collected air as compared to the headspace samples of the media and HL60 samples. The manifold, which was equipped with an Edwards Model vacuum pump and a 10,000 torr Edwards capacitance manometer, was capable of purging numerous bioreactors simultaneously. A needle valve (Swagelok, Solon, OH) and flowmeter (Dwyer Instruments Inc. Michigan City, Indiana, USA) was used to adjust the net flow to the bioreactors to 2500 cc/min. The purge time was adjusted, depending on the number of bioreactors in use, to ensure that each bioreactor was flushed with a volume of air approximately three times that of its own. After purging was completed, the stopcocks on each bioreactor were sealed at ambient pressure.


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)

The 378 mL glass bioreactor designed for incubating cells in air containing low volatile organic compounds and post incubation collection of the gaseous headspace.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Figure 1: The 378 mL glass bioreactor designed for incubating cells in air containing low volatile organic compounds and post incubation collection of the gaseous headspace.
Mentions: The Teflon vials containing the cell suspensions (40 × 106 cells/30 ml) were placed inside cylindrical glass bioreactors. The glass bioreactors were specifically designed to collect the gaseous headspace above aqueous cultures (see Figure 1) [19]. The bioreactor consisted of two glass halves joined together with an o-ring and secured by a spherical joint Thomas® pinch clamp. The bioreactor had an interior volume of 378 mL and was fitted with valves, sealed with high vacuum Chem-Vac™ stopcocks, at both ends. Once the apparatus was fully assembled it was attached to a pressurized manifold to purge the bioreactor of ambient air and replace it with air containing low levels of volatile organic compounds (VOCs) and 5% CO2. The low VOC air was prepared by doping 5% pure CO2 in to whole air collected by the Blake-Rowland lab from the rural Crooked Creek Research Station in California's White Mountains [29]. Figure 2(B) and 4(B) illustrate the low levels of selected VOCs in the collected air as compared to the headspace samples of the media and HL60 samples. The manifold, which was equipped with an Edwards Model vacuum pump and a 10,000 torr Edwards capacitance manometer, was capable of purging numerous bioreactors simultaneously. A needle valve (Swagelok, Solon, OH) and flowmeter (Dwyer Instruments Inc. Michigan City, Indiana, USA) was used to adjust the net flow to the bioreactors to 2500 cc/min. The purge time was adjusted, depending on the number of bioreactors in use, to ensure that each bioreactor was flushed with a volume of air approximately three times that of its own. After purging was completed, the stopcocks on each bioreactor were sealed at ambient pressure.

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