Dear all,
I'm currently culturing Vero cells using Eagle MEM+FBS media. A few days ago, I noticed some morphological change in my culture. The cells are getting bigger and they formed circular morphology, but still attached. Have you ever seen cell shape like this?
Hello Aqleem,
The ingredients are Eagle MEM+Penicillin+Gentamycin+Amphoterycin+L-Glutamin+FBS. Antibiotics and L-Glutamin concentration are in accordance with Sigma requirements.
HI
One possibility not mentioned is cross contamination with another cell type. This actually happens frequently. Are you working with more than one cell line in the lab? If so do you use different bottles of medium and trypsin? I would thaw a new vial of your original cell line, antibiotic free. After the first passage keep half antibiotic free and the other half on antibiotics. This should resolve the issue.
Good luck.
Hello Sreedhar,
Currently I'm just working with one cell line in my lab. So seems it's not from cross contamination.
You mentioned about the antibiotics. Is it possible the morphological change caused by antibiotics? But I've adjusted the concentration in accordance with the requirements.
Hi,
Addition of antibiotic may be killing cells, that is why they are turning round. Cells shrink, round-up and placid down when they are dying. Long term culture also may affect morphology of the cells.
what antibiotics? regular Penicillin/Streptomycin? or something else?
It is better to test and to restart culturing with a fresh stock.
Check with mycoplasma contamination.
Hope this helps little.
Hi Shreedar,
We use Penicilin+Gentamycin+Amphotericin.
Thank you for your suggestions.
Sometimes its hard to find the reason why. It may take a lot of energy to find up. I prefer to defrost a new aliquot. It is also usefull to clean everything and prepare new media.
Hi Ivana,
Yes that's what we did, finally. Thank you for your suggestion.
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