Hello. So I'm quite stumped. I have a compound I'm testing on A549 cells that results in the phenotype in the image. I have never seen this before, has anyone else? My first thought would be apoptotic blebbing, but you can see the compartments, rings, bubbles (?) are all constrained to the interior of the cells. Any help on this would be greatly appreciated!
Hi Trisha,
I have never seen a phenotype like this but it could be autophagy.
Best,
Karsten
Hi Tirsha,
These circular phenotypes looks a lot like lipid droplets. I work with myoblast cells (C2C12) and subject them to Adipgenic Differentiation and thats the same (~almost) phenotype as as the cells I work with.
Best,
Abdi
generally this cells are not suitable for continuing your work.
Hello!
Yes I have seen this phenotype previously while treating my cells with a particular inhibitor. The cells are not dead actually, they are arrested in the cell cycle and they get bigger and bigger. It was associated with a senescent state.
Djihad
Hi Trischa,
I saw similar pictures if we treat 3T3 Fibroblasten with Imidazol (and an emulgator from poloxamer typ). Imidazol are basic and I suppose thats the reason. Check if anything make your medium basic.
good luck
Jochem
Thank you all for your input! I've got a few avenues to investigate now. If I am able to pinpoint the phenotype or the cause, I'll update the post. Thanks again!
I am truly amazed by what I saw from the image you provided!
However for your information, this phenotype does exist on normal growing of the cells without any treatment, but occurs at much lower frequency.
When I noticed this, I was pretty curious and I asked my lab mate, but she said it is just dying cell. I do think that it is more than dying cell and I would love to hear more from you if you are able to investigate further.
Here I attach one of the images for untreated control group for my experimental treated group. Image was taken under 200x magnification.
I'd go with Dr Jama's interpretation. Fix the monolayers in phosphate buffered formalin and stain the cultures with Oil Red O - the droplets are lipid-filled vacuoles. Check out the mesenchymal stem cell literature for similar pics. Betcha the compound is a PPARgamma modulator or something like it.
Hi,
I had similar kind of problem and this situation is impossible to fix. I thought it was because of mycoplasma contamiation, but when tested, non of my culture had mycoplasma. I also changed the reagents, but the problem persisted. I culture mesenchymal stem cells from amniotic fluid and I still have no clear explanation for this phenotype, but this phenotype occurs mostly when the cells are thawed from cryopreservation and also when they are cultured for may passages. What you see are the vacuoles inside the cells, but I am not sure what are inside those vacuoles. The cells are not dead, but also they won't divide. They are basically at G0 phase and they can stay like it for indefinite amount of time. Notice how nucleus becomes indistinguishable, which is intriguing for me.
If you want, you can leave them as such without paying much attention (only changing medium when it dries up) to know how longer they live like this.
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