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Beijing Da Xue Xue Bao. 2013 Jun 18;45(3):489-92.

[A new methods for determining hydrogen sulfide release in cultured cells].

[Article in Chinese]

Author information

1
International Medical Center, PLA General Hospital, Beijing, China.

Abstract

OBJECTIVE:

To establish a simple method of measuring hydrogen sulfide (H2S) in cultured living cells.

METHODS:

Filtration membrane was stuck on the lid of cell culture plate. H2S released from cultured cells was trapped by zinc acetate to generate ZnS deposition. Then the ZnS trapped in the filtration membrane was measured by methylene blue assay and the H2S production from the living cells was counted according to the standard curve. This simple method was used to access the H2S release in HepG2 (high expression CBS and CSE) and HUVEC (low expression CSE) cell lines.

RESULTS:

H2S generation in cultured HepG2 cells assayed using the present method was (859.39±19.12) nmol/(min×10(6) cells). PAG (CSE inhibitor), HA (CBS inhibitor) or the two-inhibitor (PAG+HA) treatment significantly lowered H2S release, respectively: (341.34±105.90) nmol/(min×10(6) cells), (375.05±174.50) nmol/(min×10(6) cells), and (204.47±97.14) nmol/(min×10(6) cells). The H2S production of HUVEC was (26.23±3.24) nmol/(min×10(6) cells) (about 1/30 production of HepG2 cell). Trypan blue assay showed that the cell viability was greater than 95%, suggesting that there was no cytotoxicity by using the present instrument.

CONCLUSION:

The modified instrument in cell culture plate lid was feasible for detection of hydrogen sulfide release in living cells.

PMID:
23774934
[Indexed for MEDLINE]
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