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United States Department of Agriculture

Agricultural Research Service

Research Project: IMPROVEMENT OF UV RESISTANCE, VIRAL AND HOST RANGE ENHANCEMENT OF BACULOVIRUSES AS BIOCONTROL AGENTS

Location: Biological Control of Insects Research

Title: A Cell Culture Derived from the Red Flour Beetle, Tribolium castaneum

Authors
item Goodman, Cynthia
item Stanley, David
item Beeman, Richard
item Park, Yoonseong -

Submitted to: In Vitro Biology Meeting
Publication Type: Abstract Only
Publication Acceptance Date: March 10, 2010
Publication Date: December 1, 2010
Citation: Goodman, C.L., Stanley, D.W., Beeman, R.W., Park, Y. 2010. A Cell Culture Derived from the Red Flour Beetle, Tribolium castaneum [abstract]. In Vitro Biology Meeting. p. 117.

Technical Abstract: The red flour beetle, Tribolium castaneum, has become a model organism for agricultural and medical research (e.g., Li et al., 2010). We are in the process of establishing a continuously replicating T. castaneum cell line. Coupled with the recently completed genome sequence (Richards et al., 2008, Kim et al., 2010), this established line will become an important research tool. We used egg, pupa and adult stages for tissue sources. We tested numerous cell culture media (including EX-CELL 420(TM), Shields & Sang, L-15, TNM-FH, IPL-41, and Schneider’s medium with 10% fetal bovine serum and antibiotics) to determine their ability to sustain the viability and encourage the replication of T. castaneum cells. Our most promising culture was initiated by co-culturing pupal and adult tissues in EX-CELL 420 medium containing 10% FBS. We surface sterilized the beetles, minced them in medium, and then incubated the tissues in trypsin to dissociate cells. After centrifugation (800 x g for 10 minutes), we re-suspended the cells in fresh medium in T25 flasks. Thereafter, we replenished half the medium every 7-10 days. After approximately 6 months, we began completely replacing the medium weekly. This culture has now been through seven passages. We performed DNA fingerprinting using PCR on the T. castaneum cell culture and its source insect to confirm identification. The cell culture contains a variety of possible cell types, based on morphology. This represents the first report of a nearly continuously replicating cell line from T. castaneum. We hope to make this cell line available to other researchers in the near future.

Last Modified: 7/28/2014
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