title: [A new methods for determining hydrogen sulfide release in cultured cells].
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.
  
  
  CONCLUSIONS
  The modified instrument in cell culture plate lid was feasible for detection of hydrogen sulfide release in living cells.
authors.affiliation: International Medical Center, PLA General Hospital, Beijing, China.
authors.forename: Jing
authors.initials: J
authors.lastname: Xie
authors.forename: Qiang
authors.initials: Q
authors.lastname: Zeng
authors.forename: Yang
authors.initials: Y
authors.lastname: Zheng
authors.forename: Feng
authors.initials: F
authors.lastname: Liao
authors.forename: Guo-heng
authors.initials: GH
authors.lastname: Xu
authors.forename: Chao-shu
authors.initials: CS
authors.lastname: Tang
authors.forename: Bin
authors.initials: B
authors.lastname: Geng
mesh_terms: D002470:Cell Survival; D056945:Hep G2 Cells; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D006862:Hydrogen Sulfide / Q000032:analysis*
chemical_list: D006862:Hydrogen Sulfide