I am using a rather basic medium for my HEK293 cells and have seen mediocre transfection efficiency. I am wondering if I could get an increase in efficiency by simply using a richer media. If so, are there supplements you would recommend?

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Accepted Answer

It is likely not simply an issue of using a “richer” media, but more so a better (i.e. possibly different) media that allows for more rapid growth of your cells so that the highest percentage of cells as possible are actively dividing at the time of transfection to promote DNA entry into the nucleus.   There are many media developed specifically to support rapid cell growth and high density cultures and I would suggest screening some of the various 293 media available to find the one that works best for you rather than simply adding feeds or other supplements to your existing media.  Gibco FreeStyle293 media and the newer Expi293 media are specifically designed to support high density HEK293 growth and transfection.

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We are having some success in our HEK transfection but want to optimize the protocol to see if we can do better. What steps would you recommend and which do you think would have the most impact?

We have used transient production for a few projects, but are now looking at expanding our use. What kind of volumes could we reasonably expect if we scaled up and would you recommend HEK or CHO cells for high volumes? Also, if we optimized our system, what is the fastest timeline could we achieve?

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