After 24 hrs of VERO cell line thaw, we observed granules in the cell cytoplasm (attachment VERO_24hrs). The cells are in Eagle´s MEM with 10% FBS, 2 mM L-Glutamine and 50 ug/ml Gentamycin. At the same time with the same medium we thawed PK15 cells and these look fine, without any changes in cytoplasm (PK15_24hrs). I have few experiences with VERO cell line, is this normal for these cells? I don´t think it is mycoplasma, because the granules are in the cytoplasm. The black spots, which looks like in the medium are dirt on the optics :) The viability of the cells at the time of freezing (2 years ago) was 96% and the liquid nitrogen was confluently supplemented during the storage.
Does anybody expirienced this, or do you have any suggestion what may cause this?
Thank you in advance.
It is probable that what you have cultivated in vero cells (viruses or other organisms) caused the formation of stress granules which are ephemeral cytoplasmic aggregates containing stalled translation preinitiation complexes involved in mRNA storage and triage during the cellular stress response. SG formation is triggered by the phosphorylation of the alpha subunit of eIF2 (eIF2α), which provokes a dramatic blockage of protein translation. I want to know what you have grown on these cells. Look at the link below
http://www.ncbi.nlm.nih.gov/pubmed/21813702
Dear Meriem! Thank you for your answer. These cells are before infection. I thawed them for virus isolation.
I always found these cytoplasmic granule in the senescent vero cells. Just let them grow, this granule should be gone in the next passage. You will see it again, when your vero cells reach confluent and getting old.
Thank you Nithipong! I thawed cells in the 78. passage. Do you think, they are old? Should I provide new VERO cells?
Hello Everyone
I am also facing the same kind of problem in all cell lines running in our culture room. We have changed almost everything and fumigated the culture room but the problem is still persisting. Can anybody suggest, is it the contamination or anything else. I am attaching the picture
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