Submitted to: Journal of Methods of Cell Science
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/14/2001
Publication Date: N/A
Citation: N/A Interpretive Summary: Insect cell cultures are important tools in basic biology that can lead to new approaches for control of agricultural pests. This is particularly true for techniques that would be cumbersome or impossible in the whole insect. Often scientists must use an insect cell line for research that was derived from a particular pest insect, in this paper the gypsy moth. In this study we optimize the method for bringing DNA into three different gypsy moth cell lines using a technique called transfection. We determine the best gypsy moth cell line and transfection method for use in the laboratory. This information will be of interest to scientists who may need to use gypsy moth cells in their experimental work and to any industry wanting to over-express protein in gypsy moth cells.
Technical Abstract: Lepidopteran cell lines derived from the gypsy moth, Lymantria dispar, have not been widely used in protein expression studies or systems because they are weakly adherent, have specific growth requirements and characteristics, and are generally difficult to transfect. Using lipid-mediated transfection of a reporter plasmid, we improve the standard method for transfection of L. dispar-derived embryonic cell lines IPLB-LdEp and LdEIta, obtaining transfection efficiencies of 34% and 30%, respectively, as determined by image analysis assays. Using the standard lipid-mediated method, we obtain transfection efficiencies for L. dispar-derived cell line IPLB-Ld652Y of at least 41% with high mean expression levels, indicating the IPLB-Ld652Y cell line is a superior choice for expression studies or systems requiring L. dispar-derived cells.