|Christian, Peter - CSIRO-AUSTRALIA|
Submitted to: In Vitro Cellular And Developmental Biology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: August 28, 1998
Publication Date: N/A
Interpretive Summary: Insect cell lines offer an attractive, cost efficient and microbial free system for the production of baculoviruses for use as biological control agents. The conventional method of producing these agents in larvae is costly, labor intensive and the final product contains undesirable microbial contaminants. Accordingly we undertook a study to determine which of the cell lines derived from two major insect pests were the most efficient for the production of two baculoviruses that can be used as biological control agents. In the present study, several different growth media were used to established new cell lines from embryonic and ovarian tissues of host insects. Of the eight cell lines established, one cell line produced ten fold more virus than a commonly used cell line for the production of a wide host range baculovirus (AcMNPV). The second baculovirus (HzSNPV) which has a narrower host range, did not replicate to a higher level in any of the cell lines compared with a control cell line commonly used for production of this virus. This study provides new cell lines for the production of baculoviruses that can be used by industry as well as by the research community.
Technical Abstract: A total of eight cell lines were established from Helicoverpa armigera (3) and H. punctigera (5) embryos and ovaries. Cell lines were established and grown in TC100 and/or TC199-MK containing 10% fetal calf or fetal bovine serum. The serum free medium ExCellTM400 was also employed as well as supplemented with 10% fetal calf serum but failed to generate cell lines from fat body, embryos or ovarian tissues. Cell lines consisted of heterogenous cell types ranging from oval to fibroblast-like. This is the first report of the successful establishment of cell lines from H. punctigera. Cell lines from the two species were distinguishable from each other by DAF-PCR and noticeable differences in minor bands were observed amongst cell lines from the same species. All of the established cell lines from both species were susceptible to HzSNPV but did not replicate virus to a higher level than that of a H. zea cell line (BCIRL-HZ-AM1-A11). .However a H. punctigera cell line (HP1) replicated AcMNPV to the highest titer (1.0 x 10**8 TCID50/ml) and one of the H. armigera cell lines (HA1) was susceptible.