Hello Everyone,
we started working recently with the monocyte cell-line THP-1. Since it is the first time we are working with these cells, I would love to get some advise: we have started to see some round, dark shapes among the cells, and are not really sure as to what they are. These come in different sizes, and are usually sticked to a cell. They do not look like spores, fungi, or bacteria, so we are at a loss here. We have searched in the literature, and similar phenotypes seem to indicate necrosis, or necroptosis. I have attached an image, were the white arrows show the "ghosts" whithin the cell culture.
Cell culture is done in RPMI+5% FBS and 1% P/S and 37°C.
Thank you very much in advance!
Best,
Astrid
They look like blebs, herniations of the plasma membrane. These can form after enzymatic dissociation and mechanical damage in sibculturing as well as in response to physical properties of the culture medium, like hypotonicity.
https://www.nature.com/articles/cdd201081.pdf
Dear Ellen A G Chernoff
Thank you so much for your prompt response and publication. We need then to look out for elevated blebbing, and see that the cells are healthy. This will hopefully come with experience. Maybe I am treating them a little too roughly...
Dear Sudeep Kumar Maurya
Thank you for your advice! We have beginning doing that, since I read in many publications, cell culture was done with 10% FBS. So far, less blebbing has occured, so we may continue doing this.
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