Skip to main content

Perspectives on Methodology for In Vitro Culture of Helicobacter pylori

  • Protocol
  • First Online:
Helicobacter Species

Part of the book series: Methods in Molecular Biology ((MIMB,volume 921))

Abstract

Over the past 25 years, a variety of methods have been developed for culture of Helicobacter pylori in vitro. H. pylori is a capnophilic and microaerophilic organism that is typically cultured using complex culture media. Analysis of H. pylori growth in chemically defined media has provided insight into the nutritional requirements, physiology, and metabolic capacities of this organism.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Marshall BJ, Warren JR (1984) Unidentified curved bacilli in the stomach of patients with gastritis and peptic ulceration. Lancet 1:1311–1315

    Article  PubMed  CAS  Google Scholar 

  2. Ndip RN, MacKay WG, Farthing MJ, Weaver LT (2003) Culturing Helicobacter pylori from clinical specimens: review of microbiologic methods. J Pediatr Gastroenterol Nutr 36:616–622

    Article  PubMed  CAS  Google Scholar 

  3. Olivieri R, Bugnoli M, Armellini D, Bianciardi S, Rappuoli R, Bayeli PF, Abate L, Esposito E, De Aziz GLJ et al (1993) Growth of Helicobacter pylori in media containing cyclodextrins. J Clin Microbiol 31:160–162

    PubMed  CAS  Google Scholar 

  4. Westblom TU, Madan E, Midkiff BR (1991) Egg yolk emulsion agar, a new medium for the cultivation of Helicobacter pylori. J Clin Microbiol 29:819–821

    PubMed  CAS  Google Scholar 

  5. Bury-Mone S, Kaakoush NO, Asencio C, Megraud F, Thibonnier M, De Reuse H, Mendz GL (2006) Is Helicobacter pylori a true microaerophile? Helicobacter 11:296–303

    Article  PubMed  CAS  Google Scholar 

  6. Kelly DJ (2001) The physiology and metabolism of and Campylobacter jejuni and Helico­bacter pylori. Symp Ser Soc Appl Microbiol 30:16S–24S

    Google Scholar 

  7. St Maurice M, Cremades N, Croxen MA, Sisson G, Sancho J, Hoffman PS (2007) Flavodoxin:quinone reductase (FqrB): a redox partner of pyruvate:ferredoxin oxidoreductase that reversibly couples pyruvate oxidation to NADPH production in Helicobacter pylori and Campylobacter jejuni. J Bacteriol 189:4764–4773

    Article  PubMed  CAS  Google Scholar 

  8. Bury-Mone S, Mendz GL, Ball GE, Thibonnier M, Stingl K, Ecobichon C, Ave P, Huerre M, Labigne A, Thiberge JM, De Reuse H (2008) Roles of alpha and beta carbonic anhydrases of Helicobacter pylori in the urease-dependent response to acidity and in colonization of the murine gastric mucosa. Infect Immun 76:497–509

    Article  PubMed  CAS  Google Scholar 

  9. Xia HX, Keane CT, O’Morain CA (1994) Culture of Helicobacter pylori under aerobic conditions on solid media. Eur J Clin Microbiol Infect Dis 13:406–409

    Article  PubMed  CAS  Google Scholar 

  10. Cederbrant G, Kahlmeter G, Ljungh A (1992) Proposed mechanism for metronidazole resistance in Helicobacter pylori. J Antimicrob Chemother 29:115–120

    Article  PubMed  CAS  Google Scholar 

  11. Cottet S, Corthesy-Theulaz I, Spertini F, Corthesy B (2002) Microaerophilic conditions permit to mimic in vitro events occurring during in vivo Helicobacter pylori infection and to identify Rho/Ras-associated proteins in cellular signaling. J Biol Chem 277:33978–33986

    Article  PubMed  CAS  Google Scholar 

  12. Buck GE, Smith JS (1987) Medium supplementation for growth of Campylobacter pyloridis. J Clin Microbiol 25:597–599

    PubMed  CAS  Google Scholar 

  13. Hazell SL, Graham DY (1990) Unsaturated fatty acids and viability of Helicobacter (Campylobacter) pylori. J Clin Microbiol 28:1060–1061

    PubMed  CAS  Google Scholar 

  14. Khulusi S, Ahmed HA, Patel P, Mendall MA, Northfield TC (1995) The effects of unsaturated fatty acids on Helicobacter pylori in vitro. J Med Microbiol 42:276–282

    Article  PubMed  CAS  Google Scholar 

  15. Marchini A, Massari P, Manetti R, Olivieri R (1994) Optimized conditions for the fermentation of Helicobacter pylori and production of vacuolating cytotoxin. FEMS Microbiol Lett 124:55–59

    Article  PubMed  CAS  Google Scholar 

  16. Deshpande M, Calenoff E, Daniels L (1995) Rapid large-scale growth of Helicobacter pylori in flasks and fermentors. Appl Environ Microbiol 61:2431–2435

    PubMed  CAS  Google Scholar 

  17. Reynolds DJ, Penn CW (1994) Characteristics of Helicobacter pylori growth in a defined medium and determination of its amino acid requirements. Microbiology 140:2649–2656

    Article  PubMed  CAS  Google Scholar 

  18. Nedenskov P (1994) Nutritional requirements for growth of Helicobacter pylori. Appl Environ Microbiol 60:3450–3453

    PubMed  CAS  Google Scholar 

  19. Albertson N, Wenngren I, Sjostrom JE (1998) Growth and survival of Helicobacter pylori in defined medium and susceptibility to Brij 78. J Clin Microbiol 36:1232–1235

    PubMed  CAS  Google Scholar 

  20. Testerman TL, McGee DJ, Mobley HL (2001) Helicobacter pylori growth and urease detection in the chemically defined medium Ham’s F-12 nutrient mixture. J Clin Microbiol 39:3842–3850

    Article  PubMed  CAS  Google Scholar 

  21. Testerman TL, Conn PB, Mobley HL, McGee DJ (2006) Nutritional requirements and antibiotic resistance patterns of Helicobacter species in chemically defined media. J Clin Microbiol 44:1650–1658

    Article  PubMed  CAS  Google Scholar 

  22. Weinberg MV, Maier RJ (2007) Peptide transport in Helicobacter pylori: roles of dpp and opp systems and evidence for additional peptide transporters. J Bacteriol 189:3392–3402

    Article  PubMed  CAS  Google Scholar 

  23. Doherty NC, Tobias A, Watson S, Atherton JC (2009) The effect of the human gut-signalling hormone, norepinephrine, on the growth of the gastric pathogen Helicobacter pylori. Helicobacter 14:223–230

    Article  PubMed  CAS  Google Scholar 

  24. Schraw W, McClain MS, Cover TL (1999) Kinetics and mechanisms of extracellular protein release by Helicobacter pylori. Infect Immun 67:5247–5252

    PubMed  CAS  Google Scholar 

  25. Smith TG, Lim JM, Weinberg MV, Wells L, Hoover TR (2007) Direct analysis of the extracellular proteome from two strains of Helicobacter pylori. Proteomics 7:2240–2245

    Article  PubMed  CAS  Google Scholar 

  26. Schilling CH, Covert MW, Famili I, Church GM, Edwards JS, Palsson BO (2002) Genome-scale metabolic model of Helicobacter pylori 26695. J Bacteriol 184:4582–4593

    Article  PubMed  CAS  Google Scholar 

  27. Doig P, de Jonge BL, Alm RA, Brown ED, Uria-Nickelsen M, Noonan B, Mills SD, Tummino P, Carmel G, Guild BC, Moir DT, Vovis GF, Trust TJ (1999) Helicobacter pylori physiology predicted from genomic comparison of two strains. Microbiol Mol Biol Rev 63:675–707

    PubMed  CAS  Google Scholar 

  28. Kavermann H, Burns BP, Angermuller K, Odenbreit S, Fischer W, Melchers K, Haas R (2003) Identification and characterization of Helicobacter pylori genes essential for gastric colonization. J Exp Med 197:813–822

    Article  PubMed  CAS  Google Scholar 

  29. Eaton KA, Brooks CL, Morgan DR, Krakowka S (1991) Essential role of urease in pathogenesis of gastritis induced by Helicobacter pylori in gnotobiotic piglets. Infect Immun 59:2470–2475

    PubMed  CAS  Google Scholar 

  30. Tsuda M, Karita M, Morshed MG, Okita K, Nakazawa T (1994) A urease-negative mutant of Helicobacter pylori constructed by allelic exchange mutagenesis lacks the ability to colonize the nude mouse stomach. Infect Immun 62:3586–3589

    PubMed  CAS  Google Scholar 

  31. Senkovich O, Ceaser S, McGee DJ, Testerman TL (2010) Unique host iron utilization mechanisms of Helicobacter pylori revealed with iron-deficient chemically defined media. Infect Immun 78:1841–1849

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

Supported by National Institutes of Health (R01 AI039657, R01 AI068009, P01 CA116087) and Department of Veterans Affairs.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Timothy L. Cover .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2012 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Cover, T.L. (2012). Perspectives on Methodology for In Vitro Culture of Helicobacter pylori . In: Houghton, J. (eds) Helicobacter Species. Methods in Molecular Biology, vol 921. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-005-2_3

Download citation

  • DOI: https://doi.org/10.1007/978-1-62703-005-2_3

  • Published:

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-004-5

  • Online ISBN: 978-1-62703-005-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics