These are AGS cells (Gastric cancer cell line) after incubation in a low pH stale RPMI + 1% L-glutamine +10% FBS + 2% antibiotic for 10 days without medium refresh. When I check the cells most were dead and these giant ball shape structures were observed in several pieces attached to the bottom of the flask. I am sure there is no contamination, because after medium refresh and improving the culture condition these structures have disappeared quickly and cells were returned to normal. What were they? :|
Multinucleated giant cells, usually considered as result of cell fusion. Of course, mitosis not followed by cytokinesis may be possible, too. Can be stained nicely. And seem to be quite vital. But they get lost during passage. Found this in culture from avian embryonic liver, e.g. after carcinogen treatment (but in controls as well, to a lesser extent) and with BT549.
Stain the cells with nuclear dye and confirm if its a multinucleated cell.
This looks like the results of necrosis to me with the enlarged cell size plus chaotic/broken cell contents. You can google image for necrotic cells, your image looks like those. Based on my past experience, I was able to see the multiple nuclei even under the bright field, but I couldn't see them from your image. And without changing medium for 10 days...it is possible that it's under deprivation of nutrients or energy. Of course, it could also caused by multinucleation, but it happens only when you use something to induce it to happen or it's not a very stably established cells line. Flow cytometric analysis can potentially distinguish multinucleated cells from apoptotic cells or necrotic cells at the same time. You just need to stain cell with DNA maker (PI) and cell death marker and look at the distribution profile. Another possibility I can think of is senescence. It's relatively unlikely, but the large size just makes me think about it. For this case you can use B-galactosidase assay to check it.
Hope it helps!
These are genomically unstable cells with fragmented nuclei often appear as multinucleated cells. It is a response to stress (here it is nutrient depriviation), any stress that can disrupt cytoskeleton (example DMSO shock) can lead to increase in number of such giant cells (after withdrawing the stress or during the prolonged stress). Its widely considered as an in vitro phenomenon, although in vivo, similar giant cells were observed.
Under stress condition, necrotic cells or any cell may get involved in few processes, like Proliferation, Differentiation, Senescence, Mitotic cell death, Apoptosis and Necrosis. These structures are the result of necrosis.
from my experiences I think this is a waste product, dead cells and necrotic cells which is disappeared after refreshment. it is too long to leave the medium without change for 10 days
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