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the cancer cell growth - Problem with the cancer cell growth (Dec/01/2004 )

I exchanged the medium for A-549 cells 3 days before. At that time the cells were at phase of growth slightly lesser to confluent growth. When i observed the cells after 3 days of incubation at 37 C , i found all the cells were floating. I had to considered them as dead cells.
Do the cells die because of the overgrowth or was there any contamination? I am confused. Can anybody found where i am mistaken?

-Deepak Joshi-

I think it must be contamination played tricks.


I have seen cells that become overconfluent die and lift off like that. Esp. aggressive cell lines like B16.


I am not sure if this is relevant to cell line you are using but i know that for 3t3 cancer cells, once confluency is reached the cells will shortly after (few days or more) start to peel off in a sheet like formation. The cells lose their matrix attachments and instead promote cell-cell interactions as they are so closely packed now.

As long as you catch the sheet peeling off shortly after its happened (i.e. the same day or next morn for example) the cells should be able to be salvaged by spinning them down and reseeding them in fresh medium (change after 1st day for extra help) at desired confluency.

Thats really the problem with confluent in cancer cells, there are transformed in most cases so they do not reach contact inhibition when a monolayer is reached and instead strart to build up and eventually migrate from the extracellular matrix underneath.

Hope this helps.