I gonna try PEI-based transient gene expression in insect cells (High Five), the protocol described in:
Article A simple plasmid-based transient gene expression method usin...
Authors successfully used Sf900 II SFM, the transfection efficiency was as high as 90%.I wonder whether other SFMs like Ex-Cell 405 or ESF 921 support comparable transfection level? Is it wise to add preformed PEI : DNA complexes directly to HighFives cultivated in these media?
Thank you in advance,
Stan
I do not have an answer, but was wondering if you have subsequently tried to do this. We are planning to try with the ESF 921 (there was a paper using Ex-cell 420 so it likely works with Ex-Cell 405)
Hi, John
Yes, we have tried both media. At least with GFP we had about 50% (maybe more..) transfection efficiency on day 2. The major problem was strong cell clumping in ESF921; nevertheless, these aggregated cells expressed even more GFP than ones in ExCell405. We also tried other constructs with different proteins; they've worked in both media, as well as GFP.
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