wdegrip at oase.uci.kun.nl
Thu Nov 24 17:21:39 EST 2005
On Thu, 24 Nov 2005, Allison wrote:
> I am starting to culture Sf9 cells and am growing them in Grace's Insect
> Medium, supplemented, +10%FCS + gentamycin. According to invitrogen it
> should be possible to slough the cells off the plastic when it's time to
> passage them. But the cells I have stick like crazy and do not want to
> come off that way. Tapping the flask does not help. I've tried
> scraping them off gently but the viablity (by trypan blue) is not great
> 65-70%). I would like better viability before going on to the virus
> infection step.
> Any suggestions?
In our hands trypsin does not help in detaching the cells. Usually Sf9
cells adhere much less strongly in serum-free media (SF-900, InsectXpress
etc) and grow just as well. Another option is to try another supplier of
culture flasks, since the type/production of the plastic may seems to
have quiite some effect. We use Greiner flasks and cells can be quite
detached in serum-supplemented media by sloughing off.
Willem J. DeGrip
Dept. Biochemistry, UMC-286
Nijmegen Centre for Molecular Life Sciences
Radboud University Nijmegen Medical Centre
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