LEPIDOPTERAN CELL LINES AFTER LONG-TERM CULTURE IN A COMMERCIAL SERUM-FREE MEDIUM: COMPARISON OF GROWTH RATES AND BACULOVIRUS REPLICATION.

Authors: Lynn, Dwight

Submitted to: Society for Invertebrate Pathology Annual Meeting
Publication Type: Abstract Only
Publication Acceptance Date: 6/22/2004
Publication Date: 7/26/2004
Citation: Lynn, D.E. 2004. lepidopteran cell lines after long-term culture in a commercial serum-free medium: comparison of growth rates and baculovirus replication.. Society for Invertebrate Pathology Annual Meeting.

Interpretive Summary:

Technical Abstract: Several manufacturers of cell culture media began marketing serum free medium (SFM) formulations for the growth of lepidopteran cells in the 1980's to capture the potentially lucrative market of using insect cell lines for production of proteins by the baculovirus expression vector. In a few cases, cell lines have been maintained on both SFM and serum-supplemented modified TC-100 (which is a modification of Grace's medium). Cells grown in each medium were tested for susceptibility to and productivity of various multiply-embedded nucleopolyhedroviruses (MNPVs). The three lines chosen for these experiments fall into three categories of relative growth in SFM vs. TC100. LdFB cells grew similarly in each medium; LdEIta grew better in Ex-Cell than in TC-100; while Ag286 grew better in TC-100 than in Ex-Cell. Even though disparity exists in growth rates between the two media for the different cell lines, endpoint assays suggest that cells grown in serum-containing medium are more susceptible to virus infect than their SFM counterparts. Alternatively, optimal virus productivity was consistently higher (15-30%) in each line that had been grown in SFM compared with the same cells in TC-100. The virus productivity results are consistent with an earlier study in my lab in which I compared virus replication in long- and short-term passage of the LdEIta and IPLB-Sf-21 cell lines, although in that case, both of the cell lines that were tested grow faster in SFM than in serum-containing medium. The contradictory nature of the results on susceptibility and productivity may simply indicate that, while it takes greater amounts of input virus to initiate an infection in the SFM-adapted cells, once a cell is infected it appears to produce more viral occlusion bodies.