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United States Department of Agriculture

Agricultural Research Service

Research Project: EICOSANOID-MEDIATED AND MOLECULAR IMMUNE SIGNALING INHIBITORS IN PIERCING/SUCKING INSECT PESTS OF SMALL AND URBAN VEGETABLE FARMS

Location: Biological Control of Insects Research

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

Authors
item GOODMAN, CYNTHIA
item STANLEY, DAVID
item RINGBAUER, JOSEPH
item Beeman, Richard
item Silver, Kristopher
item Park, Yoonseong -

Submitted to: In Vitro Cellular and Developmental Biology - Animals
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: June 12, 2012
Publication Date: June 30, 2012
Citation: Goodman, C.L., Stanley, D.W., Ringbauer Jr, J.A., Beeman, R.W., Silver, K.S., Park, Y. 2012. A cell line derived from the red flour beetle Tribolium castaneum (Coleoptera: Tenebrionidae). In Vitro Cellular and Developmental Biology - Animals. 48:426-433.

Interpretive Summary: Insect cell lines are important tools in biomedical industries, used to produce proteins that cannot otherwise be made and used in researching insect cell-virus interactions and biological questions about insects. Many cell lines have been established from moth and fly species and these are contemporary research tools. Very few cell lines have been established from beetles, and in particular none from the red flour beetle. This is a serious shortcoming because the red flour beetle is a serious pest of stored grain and at the same time a model insect for a very wide range of research. To address this problem, we established a continuously replicating cell line from the red flour beetle. This cell line has already been provided to active researchers in ARS and other institutions, where it will benefit researchers working to devise novel approaches to controlling stored grain pests. Ultimately, this new research tool will benefit all consumers of grain-based products.

Technical 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. We characterized this line using DNA fingerprinting (DAF-PCR) and compared it with three other coleopteran cell lines and its conspecific pupae to confirm identification. 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 deg C for one hour with no decrease in viability. We recorded increased levels of one heat shock protein (43 kd) and of the hsp68 transcript following exposure to 40 deg 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.

Last Modified: 9/10/2014
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