Sf9 cells

[Grip]

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
well
detached in serum-supplemented media by sloughing off.

Wim



==============================================================================
	Willem J. DeGrip
	Dept. Biochemistry, UMC-286
	Nijmegen Centre for Molecular Life Sciences
	Radboud University Nijmegen Medical Centre
	The Netherlands
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