Freezing and thawing of cells
Posted 03 September 2010 - 07:22 AM
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Posted 04 September 2010 - 05:16 AM
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?
Posted 07 January 2012 - 09:13 AM
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.