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Treating suspension cells

cell culture suspension

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#1 doxorubicin

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Posted 25 February 2014 - 01:28 AM

Very basic question here. I normally work with adherent cells, which I culture in 10cm dishes and then make lysates by scraping cells into lysis buffer. Now I'm doing some simple experiments with cells that grow in suspension. I want to starve them for 2 hrs and then subject them to 5 different treatments to study signaling. Is it best to do this directly in small culture flasks or in falcon tubes or eppendorfs or what? I've noticed that during the 2 hr serum starvation, some of the cells start to stick to the plastic of the culture flasks, so I lose some of the cells, and if I culture the cells in falcon tubes they tend to fall down to the bottom of the cone, which will probably change my results. I guess I could put them in small dishes and scrape the cells that attach prior to collecting the suspension cells. What's the standard technique?



#2 tkf

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Posted 25 February 2014 - 12:00 PM

I would culture the cells in flasks. Any treatment that requires replacing the media can be done by spinning down the cells at 300g in conicals. Cells sticking to the flask can be removed with gentle pipetting.



#3 Tabaluga

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Posted 25 February 2014 - 01:27 PM

Agree with the above post, this is standard. Of course you should consider whether you do actually want to include those cells that settle down (vitality ?), depending on what cells you work with and what the question is.


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