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Title: Establishment of a Monarch Butterfly (Danaus plexippus, Lepidoptera: Danaidae) Cell Line and its Susceptibility to Insect Viruses

item McIntosh, Arthur
item Grasela, James

Submitted to: Journal of Applied Entomology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 1/15/2009
Publication Date: 6/9/2009
Citation: Mcintosh, A.H., Grasela, J.J. 2009. Establishment of a Monarch Butterfly (Danaus plexippus, Lepidoptera: Danaidae) Cell Line and its Susceptibility to Insect Viruses. Applied Entomology and Zoology. 44(2):331-336.

Interpretive Summary: Insect cell lines are cultures which are derived from various insect tissues and that are propagated in flasks under sterile conditions. They are invaluable in research for studying a range of parameters such as the replication of viruses, effect of toxins on such cells and gene expression. The advantages of cell cultures are that they can be grown aseptically free from contamination with bacteria and fungi, can be passaged in culture indefinitely, are easier to maintain than whole insects and can be preserved in liquid nitrogen. In the present investigation it was shown that the cells derived from this insect supported the replication of some insect viruses to very high concentrations. Since most insect cell cultures are derived from insect pests the present cell line represents a unique source coming from a non-insect pest and is the first report on its susceptibility to insect viruses. It will provide an important tool for researchers in the field of biology.

Technical Abstract: A cell line from the monarch butterfly Danaus plexippus designated BCIRL-DP-AM/JG was established from adult ovaries. The cell line consisted mainly of round cells and took a prolonged period of time in the growth medium ExCell 401 containing 10% fetal bovine serum and antibiotics before it could be subcultured on a regular basis. The cell line had a population doubling time of 32h and was susceptible to four baculoviruses (MNPV) but was refractile to a fifth baculovirus, a single nucleopolyhedrovirus (HzSNPV). PxMNPV was the most infectious of the MNPV for the monarch butterfly cell line producing a titer of 5.9 x 10**7 TCID50/ml. The non-occluded Hz-1 virus was also infectious for the D. plexippus cell line. The occlusion bodies produced by the BCIRL-DP-AM/JG cell line were infectious for 24h old T. ni larvae and gave LC50 values equivalent to or better than the LC50 for OB produced in the other susceptible cell lines by the MNPV under study. The identity of the monarch butterfly cell line was established by DAF-PCR.