HEK293 culturing

Hiromi Tagoh htagoh at hotmail.com
Tue Jan 18 19:46:58 EST 2000


"Jordi Perez Tur" <jpereztur at ibv.csic.es> wrote in message
news:38849E9A.CADB1537 at ibv.csic.es...
> Thanks a lot for your help Anna,
>
> > All that aside, it still might be possible to grow HEK293s without
> > serum.  What method have you tried for adapting the cells to the
> > serum-free conditions?  Have you tried any of the serum-substitutes
> > which are available (both Gibco and Sigma make several differnt kinds)?
> >
>
> We are trying to avoid, as much as we could, the use of any supplement
> (clearly misguided by ATCC's claims that the cells could grow without
> serum nor supplements...) because that way the purification of the
> protein (for which there is no antibody available) would be easier (we
> would monitor that the protein we have is really the one we want by
> sequencing).  It seems to me that this would not be the case, thus I
> think we will rethink the strategy.
>
>
> Thanks a lot, again, for helping us to avoid one of those dark holes....
>
> Jordi

There are several media which doesn't need adding sera to maintain the
cells.  Of course it depends on cells, but you can use some of them.  And I
don't think HEK293 require rich media.
To produce monoclonal antibodies, we sometimes use serum-free media, even
though hybridomas require rich media. In this case, we maintain cells in
media with sera, and to collect antibodies, remove sera and culture for
several days.
And also, to avoid the accumulation of metabolic waste, it would be better
to suppress the cell grow.  To decrease the serum concentlation is one way
for this.





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