Skip to main content

Advertisement

SpringerLink
Log in

A cell line derived from the red flour beetle Tribolium castaneum (Coleoptera: Tenebrionidae)

  • Published:
In Vitro Cellular & Developmental Biology - Animal Aims and scope Submit manuscript

Abstract

The red flour beetle, Tribolium castaneum, is a model organism for agricultural and medical research and its complete genome is sequenced. We established a continuously replicating T. castaneum cell line to complement existing physiological, genetic, and genomic research tools. We set up trial cell cultures from egg, pupa, and adult stages as tissue sources and incubated them in six separate cell culture media to determine the optimal combination of tissue source and medium for cell replication. Our most promising culture was generated by co-culturing adult (∼75 %) and pupal tissues in EX-CELL 420 medium containing 9 % FBS. Our new cell culture is designated BCIRL-TcA-CLG1 (TcA) and it has been subcultured more than 90 times. Amplification of genomic DNA with species-specific primers yielded DNA fragments of the expected sizes and with sequences identical to those from the published Tribolium genome. Additionally, we characterized this line using DNA fingerprinting (DAF-PCR) and compared it with three other coleopteran cell lines and its conspecific pupae to confirm identity. Its doubling time is 155.2 hr. Early passages consisted of attached cells and vesicles in suspension, whereas later passages consisted primarily of attached, spherical cells. Similar to other established cell lines, the ploidy of TcA cells was variable, ranging from 20 chromosomes/cell (diploid) to above 30 chromosomes/cell. TcA cells withstood incubation at 40°C for 1 h with no decrease in viability. We recorded increased levels of one heat shock protein (43 kDa) and of the hsp68a transcript following exposure to 40°C. Taken together, this represents the first report of a continuously replicating T. castaneum cell line. We expect the BCIRL-TcA-CLG1 line will become a useful tool in Tribolium research.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1.
Figure 2.
Figure 3.
Figure 4.
Figure 5.
Figure 6.

Similar content being viewed by others

References

  • Arakane Y.; Hogenkamp D. G.; Zhu Y. C.; Kramer K. J.; Specht C. A.; Beeman R. W.; Kanost M. R.; Muthukrishnan S. Characterization of two chitin synthase genes of the red flour beetle, Tribolium castaneum, and alternate exon usage in one of the genes during development. Insect Biochem. Molec. Biol. 34: 291–304; 2004.

    Article  CAS  Google Scholar 

  • Asgari S.; Johnson K. N. Insect virology. Caister Academic Press, Norfolk; 2010.

    Google Scholar 

  • Beeman R. W.; Stuart J. J. A gene for lindane + cyclodiene resistance in the red flour beetle (Coleoptera: Tenebrionidae). J. Econ. Entomol. 83(5): 1745–1751; 1990.

    CAS  Google Scholar 

  • Begum K.; Li B.; Beeman R. W.; Park Y. Functions of ion transport peptide and ion transport peptide-like in the red flour beetle Tribolium castaneum. Insect Biochem. Molec. Biol. 39: 717–725; 2009.

    Article  CAS  Google Scholar 

  • Charpentier C.; Tian L.; Cossette J.; Lery X. Characterization of cell lines developed from the Colorado potato beetle, Leptinotarsa decemlineata Say (Coleoptera: Chrysomelidae). In Vitro Cell Dev Biol — Animal 38: 73–78; 2002.

    Article  CAS  Google Scholar 

  • Crawford A. M. A coleopteran cell line derived from Heteronychus arator (Coleoptera: Scarabaeidae). In Vitro 18(10): 813–816; 1982.

    Article  Google Scholar 

  • Denell R. Establishment of Tribolium as a genetic model system and its early contributions to evo-devo. Genetics 180: 1779–1786; 2008.

    Article  PubMed  Google Scholar 

  • Ding L.; Candido E. P. M. HSP43, a small heat-shock protein localized to specific cells of the vulva and spermatheca in the nematode Caenorhabditis elegans. Biochem. J. 349: 409–412; 2000.

    Article  PubMed  CAS  Google Scholar 

  • Haliscak J. P.; Beeman R. W. Status of malathion resistance in 5 genera of beetles infesting farm-stored corn, wheat, and oats in the United States. J. Econ. Entomol. 76(4): 717–722; 1983.

    CAS  Google Scholar 

  • Hashimoto Y.; Zhang S.; Blissard G. W. Ao3, a new cell line from eggs of the black witch moth, Ascalapha odorata (Lepidoptera: Noctuidae), is permissive for AcMNPV infection and produces high levels of recombinant proteins. BMC Biotechnol 10(50): 1–16; 2010.

    Google Scholar 

  • Hoshino K.; Hirose M.; Iwabuchi K. A new insect cell line from the longicorn beetle Plagionotus christophi (Coleoptera: Cerambycidae). In Vitro Cell Dev Biol — Animal 45: 19–22; 2008.

    Article  Google Scholar 

  • Iwabuchi K. An established cell line from the beetle, Xylotrechus pyrrhoderus (Coleoptera: Cerambycidae). In Vitro Cell Dev Biol – Animal 35: 612–615; 1999.

    Article  CAS  Google Scholar 

  • Kim H. S.; Murphy T.; Xia J.; Caragea D.; Park Y.; Beeman R.; Lorenzen M. D.; Butcher S.; Manak J. R.; Brown S. J. BeetleBase in 2010: revisions to provide comprehensive genomic information for Tribolium castaneum. Nucleic Acids Res. 38: D437–D442; 2010.

    Article  PubMed  CAS  Google Scholar 

  • Knorr E.; Schmidtberg H.; Vilcinskas A.; Altincicek B. MMPs regulate both development and immunity in the Tribolium model insect. PLoS One 4(3, e4751): 1–14; 2009.

    Article  Google Scholar 

  • Li B.; Predel R.; Neupert S.; Hauser F.; Tanaka Y.; Cazzamali G.; Williamson M.; Arakane Y.; Verleyen P.; Schoofs L.; Schachtner J.; Grimmelikhuijzen C. J. P.; Park Y. Genomics, transcriptomics, and peptidomics of neuropeptides and protein hormones in the red flour beetle Tribolium castaneum. Genome. Res. 18: 113–122; 2008.

    Article  PubMed  CAS  Google Scholar 

  • Livak K. J.; Schmittgen T. D. Analysis of relative gene expression data using real-time quantitative PCR and 2-∆∆CT method. Methods 25: 402–408; 2001.

    Article  PubMed  CAS  Google Scholar 

  • Long S. H.; McIntosh A. H.; Grasela J. J.; Goodman C. L. The establishment of a Colorado potato beetle (Coleoptera: Chrysomelidae) pupal cell line. App. Entomol. Zool. 37: 447–450; 2002.

    Article  Google Scholar 

  • Lynn D. E. Novel techniques to establish new insect cell lines. In Vitro Cell Dev Biol – Animal 37: 319–321; 2001.

    CAS  Google Scholar 

  • Lynn D. E. Available lepidopteran insect cell lines. In: Murhammer D. W. (ed) Methods in molecular biology series. Baculovirus and insect cell expression protocols. Springer, New York, pp 117–144; 2007.

    Chapter  Google Scholar 

  • Lynn D. E.; Stoppleworth A. Established cell lines from the beetle, Diabrotica undecimpunctata (Coleoptera: Chrysomelidae). In Vitro 20(5): 365–368; 1984.

    Article  CAS  Google Scholar 

  • Mahroof R.; Zhu K. Y.; Neven L.; Subramanyam B.; Bai J. Expression patterns of three heat shock protein 70 genes among developmental stages of the red four beetle, Tribolium castaneum (Coleoptera: Tenebrionidae). Comp. Biochem. Physiol. Part A 141: 247–256; 2005a.

    Article  Google Scholar 

  • Mahroof R.; Zhu K. Y.; Subramanyam B. Changes in expression of heat shock proteins in Tribolium castaneum (Coleoptera: Tenebrionidae) in relation to developmental stage, exposure time, and temperature. Ann. Entomol. Soc. Amer. 98: 100–107; 2005b.

    Article  CAS  Google Scholar 

  • McIntosh A.; Grasela J. J.; Matteria R. L. Identification of insect cell lines by DNA amplification fingerprinting (DAF). Insect Molec. Biol. 5(3): 187–195; 1996.

    Article  CAS  Google Scholar 

  • Mitsuhashi J. Invertebrate tissue culture. Springer, Tokyo; 2002.

    Book  Google Scholar 

  • Mitsuhashi J. A. Continuous cell line from pupal ovaries of the common cutworm, Spodoptera litura (Lepidoptera: Noctuidae). Appl. Entomol. Zool. 30(1): 75–82; 1995.

    Google Scholar 

  • Park Y.; Aikins J.; Wang L. J.; Beeman R. W.; Oppert B.; Lord J. C.; Brown S. J.; Lorenzen M. D.; Richards S.; Weinstock G. M.; Gibbs R. A. Analysis of transcriptome data in the red flour beetle, Tribolium castaneum. Insect Biochem Molec Biol 38: 380–386; 2008.

    Article  CAS  Google Scholar 

  • Peel A. D. Forward genetics in Tribolium castaneum: opening new avenues of research in arthropod biology. J Biology 8(106): 1–4; 2009.

    Google Scholar 

  • Schneider I. Cell lines derived from late embryonic stages of Drosophila melanogaster. Embryo. ex. Morph. 27(2): 353–36; 1972.

    CAS  Google Scholar 

  • Schröder R.; Beermann A.; Wittkopp N.; Lutz R. From development to biodiversity—Tribolium castaneum, an insect model organism for short germband development. Dev. Genes Evol. 218: 119–126; 2008.

    Article  PubMed  Google Scholar 

  • Smagghe G.; Goodman C. L.; Stanley D. Insect cell culture and applications to research and pest management. In Vitro Cell Dev Biol - Animal 45: 93–105; 2009.

    Article  Google Scholar 

  • Stiles B.; McDonald I. C.; Gerst J. W.; Adams T. S.; Newman S. M. Initiation and characterization of five embryonic cell lines from the cotton boll weevil Anthonomus grandis in a commercial serum-free medium. In Vitro Cell Dev Biol — Animal 28: 355–363; 1992.

    Article  Google Scholar 

  • Stuart J. J.; Mocelin G. Cytogenetics of chromosome rearrangements in Tribolium castaneum. Genome 38: 673–680; 1994.

    Article  Google Scholar 

  • Tribolium Genome Sequencing Consortium. The genome of the model beetle andn pest Tribolium castaneum. Nature 452: 949–955; 2008.

    Article  Google Scholar 

  • Zhou Z.; Evans J. D.; Lu Z.; Zhao P.; Williams M.; Sumathipala N.; Hetru C.; Hultmark D.; Jiang H. Comparative genomic analysis of the Tribolium immune system. Genome Biol. 8(R177): 1–16; 2007.

    CAS  Google Scholar 

Download references

Acknowledgments

Mention of trade names or commercial products in this article is solely for the purpose of providing specific information and does not imply recommendation or endorsement by the U.S. Department of Agriculture. All programs and services of the U.S. Department of Agriculture are offered on a nondiscriminatory basis without regard to race, color, national origin, religion, sex, age, marital status, or handicap.

Author information

Authors and Affiliations

  1. Agricultural Research Service, Biological Control of Insects Research Laboratory, USDA, 1503 S. Providence Rd., Columbia, MO, 65203, USA

    Cynthia L. Goodman, David Stanley & Joseph A. Ringbauer Jr.

  2. Agricultural Research Service, Center for Grain and Animal Health Research, USDA, 1515 College Ave., Manhattan, KS, 66502, USA

    Richard W. Beeman & Kristopher Silver

  3. Department of Entomology, Kansas State University, Manhattan, KS, 66506, USA

    Yoonseong Park

Authors
  1. Cynthia L. Goodman
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. David Stanley
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Joseph A. Ringbauer Jr.
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Richard W. Beeman
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Kristopher Silver
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Yoonseong Park
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Cynthia L. Goodman.

Additional information

Editor: T. Okamoto

Rights and permissions

Reprints and permissions

About this article

Cite this article

Goodman, C.L., Stanley, D., Ringbauer, J.A. et al. A cell line derived from the red flour beetle Tribolium castaneum (Coleoptera: Tenebrionidae). In Vitro Cell.Dev.Biol.-Animal 48, 426–433 (2012). https://doi.org/10.1007/s11626-012-9524-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11626-012-9524-x

Keywords

Search

Navigation