I prepare cells for storage according to ATCC guidelines, usually 10% DMSO and 20% FBS. Cells are in cryostorage vials intended for use in liquid nitrogen. Cells are Hek293T, HepG2, HeLa and similar immortalized lines. The stocks are prepared quickly to minimize exposure to DMSO, mixed well before freezing, and frozen slowly in an ultra low freezer. After 24-48hr some stocks were moved to liquid nitrogen, others were kept in the freezer. The liquid nitrogen stocks were viable (>90%) when thawed following 1 week in liquid nitrogen and completely dead after 3 months. The freezer stocks remain viable, so far up to 6 months.
hi
This is so weird. You can use 90% FBS + 10% DMSO
(cell culture tested DMSO ,purity>98% =9ml FBS+1ml DMSO) after the vials mixed gently , so you should put the vials in Freezing container, Nalgene® Mr. Frosty(4c) then quickly at -80 ° C for 1 day .after 24h ,transfer the vial from Mr. Frosty to nitrogen tank immediately .
>>>Regularly check the nitrogen level of the cell tanks.
good luck
That is strange. Was liquid nitrogen present all the time in the Dewar and never dried even for few hours. Was liquid nitrogen tank under your control.
You can use 10% DMSO + 50% FBS, or even 10% DMSO + 90% FBS。At the same time, you need to ensure that liquid nitrogen is sufficient to cover your cells.
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