I understand speed is of the essence and that in a simple scenario with one vial you would thaw the cells in a 37 C waterbath for ~1 min until a small ice block is left. Then transfer to a 15 mL conical tube and dropwise add 10 mL medium to the side to prevent osmotic shock. Spin and plate.
But what if you have two or three vials? Would you:
1) Thaw the multiple vials all at once and leave them on ice and dilute them one by one after they've thawed?
2) Leave the vials on ice and thaw and dilute them one by one?
3) Leave the vials in -80/dry ice and take them out and thaw them one by one?
4) Other, better suggestions.
I notice other answered questions don't specifically look at the workflow in this type of scenario.
When I thawed multiple tubes I did it in groups. Similar to option 3, but I used to thaw groups of 2 to 4 if all tubes were same cells or treated similary. If not, one by one to avoid making mistakes.
To me the question looks meaningless with the info provided. The critical information relates to what medium the cells were stored in before freezing. If there are no cryoprotectants, the ice crystal artifact per se will injure most cells. The last thing to worry about is the osmotic effect. If you are able to acquire viable cells that can be plated, are the cells merely a tiny fraction of what you have put in in the first place or they represent a significant recovery?
Get one right and the rest will follow!
I agree that freezing is more critical than thawing. However, thawing, specially when multiple tubes are required, can be problem. It is not advisable to have a complete thaw (because of the risk of the liquid stars warming with the DMSO inside), and with multiple tubes that is hard to handle. Moreover, risk of making mistakes while handling several tubes trying to hurry cannot be underestimated. If you are dealing with a cell line for starting cell culture for your experiments, an excellent thawing against a very good thawing will not make a difference. However, if you wish to evaluate viability after freezing and thawing it will be critical.
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