questions about adherent cells - they are detaching (Aug/25/2005 )

my adherent cells (mouse stroma) are detaching themselves. this had happened a couple of times. is it due to confluence? are they dead?

thanks.

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Hi,

I've heard that FCS concentration can make a difference to attachment of cells. But if they attached in the first place and then started detaching, then its unlikely (assuming you used the same media for subculturing and subsequent media changes). i've had 100% confluence with no detachment. But if the cells are dead, then most likely they will start floating (in fact, that's how we determine cell viability without having to do a trypan blue assay)

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Could be cell death. I'd consider doing a trypan blue count to determine the degree of cell death - that might point you in the right direction.

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hi

may be your flask surface is not favouring cell growth...check using flasks from a different company

better luck..
anjali

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If your cells are confluent then perhaps it would seem as though your cells are detaching since they have no more space to grow.
If they are detaching at lower confluency then try using a flask from a different company (corning ones are pretty good !! )
of course you can always coat your flasks with collagen or poly-l-lysine to promote better growth.

good luck

Bose

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