Hello, some questions..mammalian cells are usually frozen between 1 x 106 cells/mL to 1 x 107 cells/mL. do you know the best concentration/cell density to freeze differentiated stem cells?I've observed that they do not like to divide as well they do after several passages. I am wondering if the amount of cells per vial could affect on it in terms of cell death or may be they just dedifferentiate. Thank you
It varies between cell types (this applies to all cells). It would be better to standardize for the stem cells you work on, by yourself..!!
Sorry I just want to ensure about the way you made the frozen stocks. Did you freez into -80 degree circumstance first for at least 1-2 days before you put into liquid nitrogen? or sometimes we consider to use cell bunker instead of medium containing DMSO for freezing medium if the cells is valuable.
I just want to remind two general points for frozen cells: always use freezing container to achieve a rate of cooling around -1°C/minute, the optimal rate for cell preservation. When recovering the cells, don't extend the time in water-bath: right after around 80% get thawed, clean the vial and transfer the content into a falcon tube with 5ml fresh medium.
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