A gas trapping method for high-throughput metabolic experiments

被引:6
作者
Krycer, James R. [1 ]
Diskin, Ciana [1 ]
Nelson, Marin E. [1 ]
Zeng, Xiao-Yi [1 ]
Fazakerley, Daniel J. [1 ]
James, David E. [1 ,2 ]
机构
[1] Univ Sydney, Charles Perkins Ctr, Sch Life & Environm Sci, Sydney, NSW, Australia
[2] Univ Sydney, Sydney Med Sch, Sydney, NSW, Australia
基金
英国医学研究理事会;
关键词
gas trap; adipocyte; cell metabolism; carbon dioxide; hydrogen sulfide; INSULIN; PHOSPHORYLATION; TISSUES; CELLS; RATS; AKT;
D O I
10.2144/000114629
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Research into cellular metabolism has become more high-throughput, with typical cell-culture experiments being performed in multiwell plates (microplates). This format presents a challenge when trying to collect gaseous products, such as carbon dioxide (CO2), which requires a sealed environment and a vessel separate from the biological sample. To address this limitation, we developed a gas trapping protocol using perforated plastic lids in sealed cell-culture multiwell plates. We used this trap design to measure CO2 production from glucose and fatty acid metabolism, as well as hydrogen sulfide production from cysteine-treated cells. Our data clearly show that this gas trap can be applied to liquid and solid gas-collection media and can be used to study gaseous product generation by both adherent cells and cells in suspension. Since our gas traps can be adapted to multiwell plates of various sizes, they present a convenient, cost-effective solution that can accommodate the trend toward high-throughput measurements in metabolic research.
引用
收藏
页码:27 / 29
页数:3
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