Refreeze thawed cells? - (Jan/19/2015 )

Hi, I work with primary hepatocytes - these cells are post-mitotic so the cells can only be used once. The cryo preserved cells are quite expensive and often the vial contains more cells than I need. Once I thaw a vial and take out a fraction, can the remaining cells be frozen back? The media used to freeze is propriety - just adding DMSO will not work. I will try this the next time I use cells - I just thought I'd ask in case someone here has thoughts or experience. 

Thanks, 

-John. 

In general no - you can't re-freeze the cells in the same mix. The formation of ice crystals and the stress of freeze/thaw cycles is enough to kill most cells fairly quickly.

I concur with bob1. 

Proprietary media recipes are sometimes available or not that crucial, so if you want, you can still try it. You are anyway going to throw away the extra cells. You can also try incubating the cells for 24 hours before re-freezing to help them recover a bit. 

I don't see high chances of success but what do I know what goes on in hepatocyte mind? Let us know!

I don't see high chances of success but what do I know what goes on in hepatocyte mind? Let us know!

So what was the result dear Aguy?