32 replies to this topic

[Rupam]

It's not hard to revive cells from frozen stock it just you can modify your procedure a little bit you will centifuge your cells at 1000rpm and for 2-3 minute...and then you will be able to get viable cells and you can store for longer period.
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Regards
Rupam

[Emilia]

Hi,
I have a problem with my Hek 293 T cells. After thawing they do not adhere to the flask. I checked if they are viable and everythig is ok.
Could be that they were too confluent when I froze them? I hit the flask for detaching the cells not to leave the too long in trypsin. is this a problem? How long can I wait untill they detach?
Thanks,

Emilia

[unknownko]

How much cells do you freeze down? Freeze/thawing generally results in a lot of cell death. Freeze down more cells in each cryovial to advert the problem of too much cell death.

I work on breast cancer cell line MD-231 and BT-549, with a similar protocol to yours for thawing, except that I spin down at a my cells at 1300rpm for 3 mins. I usually freeze down an entire fully confluent T75 into 1 cryovial.