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Reducing cell adhesion


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#1 karabas

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Posted 03 October 2012 - 06:51 AM

Hi everyone,

In our current experiment, we treat HT-29 cells with a particular compound. The result is that ~70% of the cells in the culture reduce attachment to the flask surface (but are still sticking on). We are trying to isolate these cells in a simple manner to determine if there is any difference in gene expression via qPCR and abusing this lowered cell adhesion seems to be the theoretically simplest way. Banging the flask does not work efficiently - while it does dislodge the weakly-adherent cells, we still get about a 60-40 mixture and that's not pure enough. Are there any agents that would lower cell adhesion, but not fully block it? My hope is that after our treatment the cells of interest will simply dislodge on their own and we could collect them by spinning the media down.

Any help would be appreciated!

Best,

--- Karabas

Edited by karabas, 03 October 2012 - 06:51 AM.


#2 drwho

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Posted 03 October 2012 - 11:56 AM

One way of doing this may be to use calcium free PBS this should remove any non strongly adherent cells. I have some cells where I use PBS instead to trypsin to detach them from my flask. Might be worth a try is nice and simple!

Hope this is useful




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