The cells do not confluency after thawing. Even after 10 days, the color of the medium does not change. I change the medium, the non-adherent cells disappear and then the density in the flask decreases. It has been going on like this for 3 months. Is this an adaptation problem?
Hello Nisa,
You need to ask yourself the following questions.
1. How sure are you about your freezing protocol? Have you frozen the stock using the gradual freezing technique of (-1 degree C/min) yourself?
2. Have you checked the viability count after thawing?
3. Are you using the appropriate growth media?
4. Have you tried changing the FBS lot/make?
5. Have you noticed a similar type of problem with other cultures in the same incubator? Which would mean inappropriate gas mixture in the culture environment?
6. Are you using the tissue culture treated flasks which would provide a suitable substratum for the cells to attach and grow happily?
7. Have you observed any type of contamination which may not necessarily be noticeable?
8. Have you tried getting a frozen vial of U87-MG from another lab and culturing the cells in your lab?
Finding answers to these questions may help solve your problem.
Best.
Yes there is another cell line in the same incubator. There is no adhesion or reproduction in that cell line in the same way. Thank you for your reply. I will change the brand of FBS and try it.
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