Mr Frosty - Useful??

Ainhoa M wrote:

I would like to know if anyone has been using this device (Freezin conteiner) and if it is worth buying it in order to freeze my microencapsulated cells following a slow cooling protocol.

If there is anyone who has used or is using it, I would appreciate your opinions and I would also appreciate if you could tell me how this device works and the way it is used. Just put it in the freezer and it works?

Thanks beforehand

Ainhoa -

I have used something called freezing container and it is a container that you use to put your cryogenic vials with cells in it and keep it in the -70oC until few days and then transfer the vials into the liquid nitrogen tank. Does the container hold a little amount of alcohol at the bottom? please explain more about the container and the company you will buy from.

Hi! We routinely use a Mr Frosty (or whatever VWR calls it) for "gradient" freezing of our cells. The container has some kind of matrix which has to be charged with isopropanol and colled to 4dC overnight before use (once charged, the container can be used for 4 freezing cycles, after which it has to be recharged). Vials containing cells in cold freezing medium are transferred to a chilled Mr Frosty, then moved to a -80dC freezer for a day and then transferred to LN2. the idea is to ensure the steady ~1dC cooling rate.
If you don't want to use something like this, you can follow 4dC-30-40 mins, -20dC for 4 h, then -80dC overnight before LN2 - that works too - unless your cells are super-finicky.

Its called:
Nalgene Cryo 1dC Freezing container, Cat #5100-0001, available from VWR.

I have always put my iced cells to be frozen inside a styrofoam box (we just use the ones they ship enzymes to us in) and put that inside the -80 overnight before transferring to liq. N2. The styrofoam provides slow cooling to -80. I have not had any problems using this protocol.

macbride wrote:

I have always put my iced cells to be frozen inside a styrofoam box (we just use the ones they ship enzymes to us in) and put that inside the -80 overnight before transferring to liq. N2. The styrofoam provides slow cooling to -80. I have not had any problems using this protocol.

Styrofoam holders have always worked for me. They provide slow cooling in the -80oC freezer and you can leave them there overnight or up to 2 to 3 days and then transfer the vials to liquid nitrogen.

We have found them very useful in our labs. They give you a 1 degree per minute drop in a minus 70-80 freezer. They are a lot cheaper than the liquid nitrogen-based cell freezers and just as effective, for smaller batches of cells that is.

I have not tried to freeze encapsulated cells cells with them, but I would believe it to be one of the more gentle ways to attempt to accomplish it.

I have been using the new CoolCell product with much success.  It freezes with an almost identical profile to Mr. Frosty, but does not require isoproponal.  The foam insulation also prevents freezer-burn.
http://www.capitalbiosciences.com/product/info/coolcell-nbcs-136.html

VWR carries also Coolcell, a cell freezing container, that works with no alcohol, no maintenance, very equal freezing rate at -1C/min works for all cells

VWR CoolCell

COOLCELL -1C/MIN FREEZER 
  CAT # 95059-860
Each
$89.00

Dear Samm -

not sure you area ware that Rusty B. from scientistsolutuions actually tested CoolCell as an alternative to Mr. Frosty...see prodcut review on the bttm of the page of CoolCell

Here are the CoolCell instructions for anyone interested...

I apologize for turning this discussion into an advertisment.  I only meant to provide the link as reference.

mikeee - Thanks for the link.

We use Mr.Frosty, VWR inour lab regularly.
Biju

There seems to be plenty of good data out there on the freezing rates for all the methods talked about here and some nice testimonials for the commercial devices,  but has anyone actually quantified and compared the cell survival rates when thawing identical samples of their cell lines from each of these different device types?

1. the Mr Frosty
2. the CoolCell
3. Your vial in a sandwich of two Styropfoam racks that used to hold 15ml falcons

I have used and tested a controlled rate freezer, vs Mr. Frosty, vs stacked styrofoam 15 ml tube holder, using freshly isolated buffy coat PBMC using flow cytometry as an endpoint at a commercial enterprise.  We did not find a statistically significant difference in the three devices even though we had earmarked $25k for a controlled rate freezer.  I also attended a conference recently where a large pharma core repeated the same experiment with similar results using stable transgenic CHO cells, so I reasonably confident in the use of the free alternative to these products at least for these two cell types.