ESTABLISHMENT OF CELL LINES FROM NINE LEPIDOPTERAN SPECIES

Authors: Goodman, Cynthia; McIntosh, Arthur; EL SAYED, GALAL - UNIVERSITY OF MISSOURI; Grasela, James; STILES, BRAD - AMERICAN CYANAMID

Submitted to: In Vitro Cellular And Developmental Biology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/22/1998
Publication Date: N/A
Citation: N/A

Interpretive Summary: Scientists are always searching for new ways to control pest insects which destroy important crops. One good way to do this is through the use of insect viruses, which offer a safe and effective means of killing certain kinds of insects. The viruses we work with are called "baculoviruses." Many agrichemical companies are interested in marketing these viruses, but they don't want to produce them in living insects (the major method of production until recently). Companies would like to find an easier, less expensive way of manufacturing viruses, such as by producing them in insect cells. We have been studying ways of improving this process. One important way of improving the production of viruses by insect cells is to develop new cell cultures. Each cell culture can produce different amounts of virus. By developing new cultures, we may find some cells which can produce larger amounts of virus than previously possible. We have developed 27 new cultures to aid in this effort. These cultures were made from many different types and ages of insects. Each culture varies in how it looks and grows. Because of these and other differences, we expect the cell cultures we have developed to vary in how well they produce insect viruses. In the future, we plan to see how much virus each of these cultures can produce. Eventually, we plan to use the best cell cultures for manufacturing baculoviruses.

Technical Abstract: Insect cell lines are becoming increasingly important in the propagation of baculoviruses for use as biocontrol agents. For this reason, a study was undertaken to establish cell lines from under-represented species/tissues/ stages with the eventual goal of determining if these lines are good producers of baculoviruses. A total of 27 continuous cell lines from 9 species of lepidopteran insects was generated. These cell lines were established from a variety of tissues, including embryonic, ovarian, testes and fat body. All stages of the insect life cycle were represented by the cell lines, with most of the lines being established from eggs, larvae and adults. The cell lines vary widely in their morphology and growth characteristics, suggesting that they may respond differently to infection by baculoviruses. Additionally, many of the cell lines have been adapted to grow in serum-free medium, an important first step if they will be used in large scale virus production. Lastly, an identity for each cell line has been established using biochemical methods, specifically isoelectric focusing (for isoenzyme analysis) and PCR (for DNA fingerprinting).