I'm having a problem in maintaining H9C2 cells for a long duration. After 2-3 splits cells become unhealthy as black particles start coming. Also sometimes this problem happens from the beginning. I'm using DMEM media with high glucose, Sod. Bicarbonate 1.5g/l, glutamine 4mM, 10%FBS (GIBBCO) and all others are from SIGMA.
There are a lot of factors few which I have information may be help for you. Dark dots in cell culture may be the cell debris, if it, then the darken particle will be of differing in size. But if these dots are of same size and there is also amplification observed this indicates there must be some bacterial contamination. There is also one more another case for it, that is degranulation in cell culture due to stress in your culture which also appears as dark dots.
Yes Ravi, Can you please tell what are the factors I should take care regarding stress because temp and Co2 is always optimum, rest I have humidity factor which I have tray system. else?
Thanks for the reply Doris. if any one know what exactly H9C2 media should have because they are very sensitive.
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