Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Research
  • Published:

Large Scale Cultivation of the Free Living Nematode Caenorhabditis elegans

Bio/Technology volume 12, pages 51–54 (1994)Cite this article

Abstract

We describe conditions for reproducible large scale cultivation of the free living nematode, Caenorhabditis elegans, and downstream processing of large quantities of this tissue. We were able to grow and harvest 500 grams of purified worms from a 150–liter bioreactor. Cultivation on this scale was accomplished in three steps by using worms from 50 agar plates as inoculum for 4 liters of liquid media. Mature worms from the 4–liter culture were transferred to 20 liters of fresh liquid media. After three days of incubation, the 20–liter culture was used for inoculation of the 150–liter tank. At the time of harvest, a mixture of developmental stages of worms was present. Healthy, motile, egg–laden adults worms appeared to make up the major portion of the population. Membranes prepared from sucrose purified worms were biologically active in an ivermectin binding assay. The affinity of ivermectin (Kd) and the density of receptors (Bmax) were comparable to values generated using membranes from worms grown on agar plates. The amount of membrane tissue obtained from the 500 gm worm harvest made it feasible to consider purification to homogeneity of a nematode protein such as the ivermectin receptor, which is not abundant in crude homogenates. In addition, RNA isolated from sucrose purified worms was translated in Xenopus oocytes. Several channel proteins expressed from this RNA have been characterized, including a chloride channel sensitive to ivermectin and glutamate.

Access through your institution
Buy or subscribe

This is a preview of subscription content, access via your institution

Access options

Access through your institution

Buy this article

  • Purchase on SpringerLink
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Similar content being viewed by others

References

  1. Sulston, J. and Hodgkin, J. 1988. Methods, p. 587–606. In: The Nematode Caenorhabditis elegans. Wood, W.B. (Ed.). Cold Spring Harbor Laboratory Press, New York.

    Google Scholar 

  2. Chalfie, M. and White, J. 1988. The nervous system, p. 337–391. Ibid.

    Google Scholar 

  3. Horvitz, H.R. 1988. Genetics of cell lineage, p. 157–190. Ibid.

    Google Scholar 

  4. Coulson, A., Sulston, J., Brenner, S. and Karn, J. 1986. Toward a physical map of the genome of the nematode Caenorhabditis elegans. Proc. Natl. Acad. Sci. USA 83: 7821–7825.

    Article  CAS  Google Scholar 

  5. Coulson, A., Waterson, R., Kiff, J., Sulston, J. and Kohara, Y. 1988. Genome linking with yeast artificial chromosomes. Nature 335: 184–186.

    Article  CAS  Google Scholar 

  6. Watson, J.D. 1990. The human genome project: past, present and future. Science 248: 44–48.

    Article  CAS  Google Scholar 

  7. Lewin, R. 1990. A worm at the heart of the genome project. New Scientist August 25: 38–42.

    Google Scholar 

  8. Wood, W.B. 1988. Introduction to C. elegans biology, p. 1–16. In: The Nematode Caenorhabditis elegans. Wood, W.B. (Ed.). Cold Spring Harbor Laboratory Press, NY.

    Google Scholar 

  9. Schaeffer, J.M. and Haines, H.W. 1989. Avermectin binding in Caenorhabditis elegans: A two-state model for the avermectin binding site. Biochem. Pharm. 38: 2329–2338.

    Article  CAS  Google Scholar 

  10. Schaeffer, J.M. and Bergstorm, A.R. 1988. Identification of gamma-amino-butyric acid and its binding sites in Caenorhabditis elegans. Life Sci. 43: 1701–1706.

    Article  CAS  Google Scholar 

  11. Schaeffer, J.M., White, T., Bergstorm, A.R., Wilson, K.E. and Turner, M. J. 1990. Pesticide Biochem. and Physiol. 36: 220–228.

    Article  CAS  Google Scholar 

  12. Vanfleteran, J.R. 1976. Large scale cultivation of a free-living nematode. Experientia 32: 1087–1088.

    Article  Google Scholar 

  13. Salih, N.E. and Hasan, S.A. 1987. Effect of pH of the medium on growth and reproduction of Caenorhabditis (Nematodia: Rhabditidae). J. of Biol. Sci. Research 18(1): 137–143.

    Google Scholar 

  14. Cully, D.F. and Paress, P.S. 1991. Solubilization and characterization of a high affinity Ivermectin binding site from Caenorhabditis elegans. Molecular Pharmacology 40: 326–336.

    CAS  PubMed  Google Scholar 

  15. Rohrer, S.P., Meinke, P.T., Hayes, E.C., Mrozik, H. and Schaeffer, J.M. 1992. Photoaffinity labeling of avermectin binding sites from Caenorhabditis elegans and Drosophila melanogaster. Proc. Natl. Acad. Sci. USA 89: 4168–4172.

    Article  CAS  Google Scholar 

  16. Arena, J.P., Liu, K.K., Paress, P.S. and Cully, D.F. 1991. Avermectinsensitive chloride currents induced by Caenorhabditis elegans RNA in Xenopus oocytes. Molecular Pharmacology 40: 368–374.

    CAS  PubMed  Google Scholar 

  17. Arena, J.P., Liu, K.K., Paress, P.S., Schaeffer, J.M. and Cully, D.F. 1992. Expression of a glutamate-activated chloride current in Xenopus oocytes injected with Caenorhabditis elegans RNA: evidence for modulation by avermectin. Molecular Brain Research 15: 339–348.

    Article  CAS  Google Scholar 

  18. Buckland, B., Brix, T., Fastert, H., Gbewonyo, K., Hunt, G. and Jain, D. 1985. Fermentation exhaust gas analysis using mass spectrometry. Bio/Technology 3: 982–988.

    CAS  Google Scholar 

  19. Brix, T., Drew, S.W. and Buckland, B.C. 1988. Fermentation process development within a computer controlled pilot plant, p. 151–193. In: Computers in Fermentation Technology. Bushell, M.E. (Ed.). Elsevier Science, Amsterdam.

    Google Scholar 

  20. Lu, N.C., Cheng, A.C. and Briggs, G.M. 1983. A study of mineral requirements of Caenorhabditis elegans. Nematologia 29: 425–434.

    Article  CAS  Google Scholar 

Download references

Author information

Author notes

  1. Barry Buckland: Corresponding author.

Authors and Affiliations

  1. Merck Research Laboratories, P.O. Box 2000, Rahway, New Jersey, 07065

    Kodzo Gbewonyo, Susan P. Rohrer, Leonard Lister, Bruce Burgess, Doris Cully & Barry Buckland

Authors
  1. Kodzo Gbewonyo
    View author publications

    Search author on:PubMed Google Scholar

  2. Susan P. Rohrer
    View author publications

    Search author on:PubMed Google Scholar

  3. Leonard Lister
    View author publications

    Search author on:PubMed Google Scholar

  4. Bruce Burgess
    View author publications

    Search author on:PubMed Google Scholar

  5. Doris Cully
    View author publications

    Search author on:PubMed Google Scholar

  6. Barry Buckland
    View author publications

    Search author on:PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gbewonyo, K., Rohrer, S., Lister, L. et al. Large Scale Cultivation of the Free Living Nematode Caenorhabditis elegans. Nat Biotechnol 12, 51–54 (1994). https://doi.org/10.1038/nbt0194-51

Download citation

  • Received:

  • Accepted:

  • Issue date:

  • DOI: https://doi.org/10.1038/nbt0194-51

This article is cited by

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing