Hello every one
We had a problem with tissue culture: the cells are not proliferating, the cultured amount of cell stay the same, no growth! And there are changes in the shape of cells. We suspected contamination so we clean every thing and change the media but still the same. We suspect mycoplasma infection but the test result was negative.
Do you have any idea .what is the problem.
Thanks in advance for you help!
Definitely, vacuolization in pic 3 seems to be more intensive. If no micoplasma, try to change FBS (of different serial number). Either try to thaw another vial with culture, as soon as the primary could be freezed with mistakes
Hi,
I think you need to include more detail in your question.
What cells are you culturing? I'm guessing from the labeling on the images they are T47D, MDA and MCF7? What are your culture conditions and what is the time frame in which you are observing no cell growth? What is the shape change you have observed?
What media are you using? Are you using RPMI, IMDM, or something else? Depending on cell types possibly supplementing the media with FBS, bicarb, IL's, L-Glutamine, or something else could help growth but check with Literature first.
what type of test did you used to check contamination ? try to make a Gram stain to check bacterial contamination. some times sub culturing helps in reactivation of some slow growing cell lines. in addition, check the incubation condition..
thank you every body for all suggestions, the problem was solved!I used FBS from different serial number and the cell became good .
I suggest you take a look at this site with very detailed protocols and tips on the use of cell cultures
http://cellculture.altervista.org
hope it will be useful!
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