When I treat MDCK cells (adherent epithelial cells from dog's kidney) with different metal ions, they start to detach from the cell culture flask surface. These kinds of cells are often regarded as dead, but they do not become stained with trypan blue. If I transfer those detached cells to new culture flask with fresh medium, they remain detached and they still don't become stained with tripane blue. If we perform some cytotoxicity assay (such as MTT assay), we regard that kind of cells as dead, as they are washed away during protocol (but I've observed, that they are still metabolically active). Is it possible that those cells have become senescent? Or do you have some other possible explanation?
I would like to thank you in advance for all the answers/comments.
I agree with the answer from Attila. Many of the primary cells need contact/adherence, which provide them signalling for proliferation e.g. lymphocytes from spleen/blood, keratinocytes etc. These cell colonies are usually a bunch of cells. When they are dispersed/single, it means they are either dead or on the verge of death.
I have not worked with cell-lines, but probably the same argument works there also, as in your case.
Such cells need contact ie, adherence for proper growing. From this viewpoint floating ones are died or dieing. Loss of adherence usually initiates apoptosis, i think. It could be a longer process and you can still get metabolic activity working for apoptotic machinery. Try to wash away metals by incubating cells in some EDTA/PBS next a centrifugation. Probably your floating cell will revive and attach again.
I agree with the answer from Attila. Many of the primary cells need contact/adherence, which provide them signalling for proliferation e.g. lymphocytes from spleen/blood, keratinocytes etc. These cell colonies are usually a bunch of cells. When they are dispersed/single, it means they are either dead or on the verge of death.
I have not worked with cell-lines, but probably the same argument works there also, as in your case.
kidney cells are dependent and need attachment for growth. If we add EDTA or other chelating agents will it not reduce the cells attachment to plate and thereby reduce the growth of cells
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