Recently I have been observing some strange bloomed like structures as in the attached photo in my Hela cell culture which look so different from my long term experience of culturing Hela cells. Is it any kind of fungal contamination? Does anybody have experienced such problem before? To be noted, these Hela cells are overexpressed with the catalytic domain of Poly (ADP-ribose) polymerase (PARP) targeted to be expressed in cytosol. And they have been maintained in DMEM supplemented with 10% FBS, 1% PEN-STREP, 1% L-gln, 1% Na-Pyr and have the antibiotic concentration of 650 ug/ml geneticin G-418. The overexpression of PARP did not seem to cause any problem with their culturing before.
Those appear to be giant/polyploid cells. (see Article Multinucleated Giant Cancer Cells Produced in Response to Io...
for some examples).As I recall, these cells generally form in response to cellular stresses and exhibit much higher resistance to treatments. The role of these cells in cancer is a active field of study (Article Polyploid giant cancer cells: Unrecognized actuators of tumo...
).
Hi there Hanan Ashrafi
These cells remind me of HepG2 cells in culture with these little islands they form on flat surfaces.
In those cases, there are also some weird things similar to what you see here. In some cases its a cluster of dying cells spitting out vesicle (super bright bubble like things), while some cells which have empty space nearby, expand to form large cells either polyploid or not. These are however all normal in context of culturing cancer cells.
Hope this helps.
Best
Aparajita
Thanks John and Aparajita for sharing your knowledge. I agree with your ideas. Hence, my idea is that the stress that causing the generation of these multinucleated cells in my culture is the cyto-PARP itself, since we previously observed that the cells sometimes do not want to tolerate PARP to some point, and also the expression being lost after few passage.
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