insect cell line problems

David-Paul Minde via methods%40net.bio.net (by davidminde from gmail.com)
Mon Feb 22 10:21:13 EST 2010


... why baculovirus at all (unless you happen to receive the working
system from a colleague:) ?  there is an efficient transient
transfection protocol with PEI. Much easier, faster and fewer
variables. Yield excellent as well. Only one "pet" per protein
expression trial ;)
( Methods Mol Biol. 2009;498:199-227.
High-throughput insect cell protein expression applications.

Buchs M, Kim E, Pouliquen Y, Sachs M, Geisse S, Mahnke M, Hunt I.

Biologics Center, Novartis Institutes for Biomedical Research, Basel,
Switzerland. )
cheers,
David

2010/2/22 Carmen P Pena Diaz <C.P.Pena-Diaz from 2006.hull.ac.uk>:
>
> Dear All:
>
> We recently started tissue culture work with SF9 insect cell line for protein expression.
>
> Since we obtained the cell line we started culturing in TC100 insect media and then shifted to Grace's supplemented insect media.  After the passage the cells failed to continue growth and that was almost 3 weeks ago.  We're not keeping the cultures with antibiotics in order to help them grow better.
>
> My boss obtained a second batch of cells from the firm and put them to grow directly in Grace's media, in case the media changing might have affected the cells irreversibly.  Nevertheless, they're still not growing properly.
>
> On the other hand, we also have many cells not attaching to the surface as expected.  Since this is actually needed to calculate baculovirus titer, I have not started trasnfections due to these problems.
>
> Anyone can suggest/comment on my cell culture problem?
>
>
> thank you very much
>
>
> Priscila
>
>
>
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-- 
David Minde MSc (TUM)

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