Three days ago, i thawed Hela cancer cells from liquid nitrogen but until now they are still suspended in the media and very very low number of cells became attached to the tissue culture dish.
So, i want to know what is the problem?
The media i used is DMEM (low glucose) with 10% FBS.
Thanks in advance.
Floated cells are dead cells and reasons are many like the problem with liquid nitrogen container and cryoprotectant etc. Remove the floated cells and incubate attached cells for more time, I am sure they will take morphology and start dividing in a few days.
Hi)
HeLa are fast growing cells and one can not expect such difficulties. Could be two main reasons: (i) the original stock was bad snap-frozen and your cells are dead or (ii) you have not washed the stock from dmso on fresh FBS media. There is a little chance tgat that your cells are contaminated by micoplasma but I wouldn't worry about it so far. Just take a fresh frozen aliquot, thaw it, wash/spin 1 time in FBS medium or in PBS, resuspend in FBS medium and plate your cells.
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