title: A thin-layer liquid culture technique for the growth of Helicobacter pylori.
abstract: OBJECTIVE
  Several attempts have been successful in liquid cultivation of Helicobaccter pylori. However, there is a need to improve the growth of H. pylori in liquid media in order to get affluent growth and a simple approach for examining bacterial properties. We introduce here a thin-layer liquid culture technique for the growth of H. pylori.
  
  
  METHODS
  A thin-layer liquid culture system was established by adding liquid media to a 90-mm diameter Petri dish. Optimal conditions for bacterial growth were investigated and then viability, growth curve, and released proteins were examined.
  
  
  RESULTS
  Maximal growth of H. pylori was obtained by adding 3 mL of brucella broth supplemented with 10% horse to a Petri dish. H. pylori grew in both DMEM and RPMI-1640 supplemented with 10% fetal bovine serum and 0.5% yeast extract. Serum-free RPMI-1640 supported the growth of H. pylori when supplemented with dimethyl-beta-cyclodextrin (200 microg/mL) and 1% yeast extract. Under optimal growth, H. pylori grew exponentially for 28 hours, reaching a density of 3.4 OD(600) with a generation time of 3.3 hours. After 24 hours, cultures at a cell density of 1.0 OD(600) contained 1.3 +/- 0.1 x 10(9 )CFU/mL. gamma-Glutamyl transpeptidase, nuclease, superoxide dismutase, and urease were not detected in culture supernatants at 24 hours in thin-layer liquid culture, but were present at 48 hours, whereas alcohol dehydrogenase, alkylhydroperoxide reductase, catalase, and vacuolating cytotoxin were detected at 24 hours.
  
  
  CONCLUSIONS
  Thin-layer liquid culture technique is feasible, and can serve as a versatile liquid culture technique for investigating bacterial properties of H. pylori.
authors.affiliation: Department of Microbiology, Gyeongsang National University School of Medicine, Jinju, Gyeong-Nam 660-751, Korea.
authors.forename: Jung-Soo
authors.initials: JS
authors.lastname: Joo
authors.forename: Kyung-Chul
authors.initials: KC
authors.lastname: Park
authors.forename: Jae-Young
authors.initials: JY
authors.lastname: Song
authors.forename: Dong-Hyun
authors.initials: DH
authors.lastname: Kim
authors.forename: Kyung-Ja
authors.initials: KJ
authors.lastname: Lee
authors.forename: Young-Cheol
authors.initials: YC
authors.lastname: Kwon
authors.forename: Jung-Min
authors.initials: JM
authors.lastname: Kim
authors.forename: Kyung-Mi
authors.initials: KM
authors.lastname: Kim
authors.forename: Hee-Shang
authors.initials: HS
authors.lastname: Youn
authors.forename: Hyung-Lyun
authors.initials: HL
authors.lastname: Kang
authors.forename: Seung-Chul
authors.initials: SC
authors.lastname: Baik
authors.forename: Woo-Kon
authors.initials: WK
authors.lastname: Lee
authors.forename: Myung-Je
authors.initials: MJ
authors.lastname: Cho
authors.forename: Kwang-Ho
authors.initials: KH
authors.lastname: Rhee
mesh_terms: D003470:Culture Media / Q000378:metabolism; D046508:Culture Techniques / Q000379:methods*; D016480:Helicobacter pylori / Q000254:growth & development* / Q000378:metabolism
chemical_list: D003470:Culture Media