After several days in culture, rounded and refringent cells begin to appear. Over time, their number increases until they eventually die and detach from the flask's surface. Phase contrast images at 200x magnification at 3, 7 and 14 days in culture. All other HSF cultures in the same incubator are doing fine. Culture media controls are OK (no pH changes/ no turbidity). Culture conditions: 5% CO2 + 80% RH; MEM + 10% FBS + penicillin/streptomycin. Thank you!
Hello Victor,
It could be a virus, which you might be able to narrow down by transferring a small amount of culture supernate to one of your other cultures that does not have this problem. another possibility is mycoplasma, for which there are commercial test kits, but from the pictures you posted I doubt mycoplasma is the cause. If it does transfer to other cultures, then you might think about doing (or asking a friend to do) some elctron microscopy, which should show up a virus infection on this scale.
Cheers,
David
Dear Victor, for the mycoplasma test you could try the following protocol: http://www.lifescience.net/protocols/879/mycoplasma-test/
Protocol is simple, it is a standard PCR, you just need to order the primers (primer sequences are included in the protocol).
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