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A Thin‐Layer Liquid Culture Technique for the Growth of Helicobacter pylori
Author(s) -
Joo JungSoo,
Park KyungChul,
Song JaeYoung,
Kim DongHyun,
Lee KyungJa,
Kwon YoungCheol,
Kim JungMin,
Kim KyungMi,
Youn HeeShang,
Kang HyungLyun,
Baik SeungChul,
Lee WooKon,
Cho MyungJe,
Rhee KwangHo
Publication year - 2010
Publication title -
helicobacter
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.206
H-Index - 79
eISSN - 1523-5378
pISSN - 1083-4389
DOI - 10.1111/j.1523-5378.2010.00767.x
Subject(s) - petri dish , bacterial growth , fetal bovine serum , microbiology and biotechnology , chemistry , biology , bacteria , biochemistry , in vitro , genetics
Background and Aims: 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‐β‐cyclodextrin (200 μg/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 × 10 9 CFU/mL. γ‐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 .