I have attached two pictures of A549 cell line. In the following pic the cell have protruding of some types of fiber like substance from the cell. I think there is change in phenotype of the cell line. I am trying to isolated the exosome from the same cell line. I think this may effect the yield of my exosome. I have been through this problem from last month. Because of this I bought the new cell again from the Korean cell bank. But its showing the same type of phenomenon. I don't what kind of contamination it is. The pH of my cell media is ok and there is no development of turbidity in the media too. I am guessing its a kind of mycoplasma contamination. Can any one suggest me to solve this problem?
Mycoplasma contamination is not easy to spot under the microscopy and it doesn't make the medium turbid. What you can spot is alteration of the cell morphology due to the contamination, mainly if the culture is heavily contaminated. It seems to me that this fiber-like structure is actually a wool fiber from the pipette, but it could be a fungus hypha as well. To rule out the mycoplasma contamination, I suggest doing a quick DAPI staining. But bear in mind that this technique has low sensitivity and there are plenty of detection methods much more sensitive available comercially. Attached is a picture showing how a culture heavily contaminated with mycoplasma looks like after DAPI staining.
Good luck.
https://www.google.co.uk/search?q=dapi+staining+mycoplasma&client=firefox-b&source=lnms&tbm=isch&sa=X&ved=0ahUKEwjq-8z5g83NAhXKCMAKHQe7BlQQ_AUICCgB&biw=1280&bih=915#imgrc=dFPnK_-Xom_PgM%3A
You can use MycoAlert to test your cell line:
http://www.lonza.com/products-services/bio-research/cell-culture-products/mycoplasma-detection-and-removal/mycoalert-mycoplasma-detection-kit.aspx
If it is positive discard the cells and get a new cell line batch.
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