Cryopreservation (freezing cells) without using Freezing Container
Posted 12 February 2002 - 06:21 AM
Posted 12 February 2002 - 08:36 PM
stryofoam box in a -80 C freezer for 24 hours then transferring directly to liquid nitrogen storage. The box should already be in the freezer before you start. I've done this a couple of hundred times and always had viabilities of at least 70%.
Posted 13 February 2002 - 10:08 PM
Posted 03 August 2004 - 06:34 AM
Posted 04 August 2004 - 12:05 PM
Posted 06 August 2004 - 10:40 AM
Posted 10 August 2004 - 12:16 PM
Posted 23 October 2004 - 01:16 AM
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Posted 16 December 2004 - 02:06 PM
Posted 06 October 2009 - 03:46 PM
I agree with Daniel. Although no two cell types will react the same to cryopreservation, we also find that the 10% DMSO in FBS works much better than 10% DMSO in DMEM in almost all cell types.
I agree that the final 10%DMSO+90%FBS seems to be good for most of cell types.
To be more gentle, don't rush to mix cells with DMSO. Resuspend them first in 100%FBS and let them calm down on ice for 15-30min while you are labeling cryovials etc. Then add dropwise equal volume of 20%DMSO+80%FBS, mix well, aliquot into cryovals, and freeze them either in the expensive Mr.Frosty, or a cost-nothing foam box.
Posted 14 March 2011 - 10:41 PM
i agree with Nevermore. I worked on HepG2 cells and these cells were stored with 60% FCS + 30% DMEM + 10 % DMSO. We use cryovial from nunc and we first keep them in -20 oC (2 hr), followed by -80 oC (Overnight) and next morning we store the vial in Liquid N2 cylinder.
THis works well.
Posted 08 April 2011 - 04:59 PM
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