Hello Everyone!
I am working on U-87 MG cell lines and I follow the protocol as provided by ATCC. I have recently split the U-87 mg cells and found a cloudy appearance under 20X magnification. And under 40X magnification, I find these small irregular light structures in clusters and in linear structures. These structures were around 4 to 5 in the cell flask. Can anyone suggest, if this can be a mycoplasma contamination. The media and the cells were fine in the flask.
Furthermore, there is another flask where I have observed these clusters of cells with slight discoloration (in the other image attached). I am not sure, if there is something to worry about it.
Looking forward for solution for this problem. Appreciate your time and valuable response!
Hello Akshaya Nagarajan
It is not contamination.
From the pic it seems clear that the cells are seeded at a high density. U-87MG cells are known for their tendency to aggregate and form clusters, especially when cultured at high densities. The presence of these structures can affect the overall morphology and uniformity of the culture, potentially impacting experimental results.
To minimize the formation of these clusters, you may reduce the seeding density. You may seed U-87MG cells at a lower initial density, at 1 x 10^4 cells/cm2. During passaging, you may remove the clusters by waiting for them to settle to the bottom of the flask, and then carefully aspirating the supernatant containing single cells.
Best,
Just my educated guess, it is a mold if it increases in size over time. Mycoplama doesnt look like cloudy entity.
You cannot see mycoplasma under 40* magnification. Mycoplasma staining with DAPI appears as a fine particulate or filamentous staining over the cytoplasm at 100X magnification... so educated guess would be yeast... If you are concern about mycoplasma, you can try this very simple and straightfoward product : https://www.invivogen.com/mycostrip
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