Cryopreservation - Encapsulated Cells

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Ainhoa M
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Cryopreservation - Encapsulated Cells

Post by Ainhoa M » Fri Jan 13, 2006 3:35 pm

Hi all!
I am trying several cryopreservation protocols to cryopreserve encapsulated cells (C2C12 mioblasts)but my cell viability results after thawing aren't as good as previous to cryopreservation. In my first trials I used 10%DMSO but after having a 40% decrease in cell viability, I tried higher percentages of DMSO, with even worse results. I am trying 5% and 1% DMSO next week but I am not sure this might be the solution.
I also tried different freezing protocols, such as: 1h at -20ºC, 23h at -80ºC followed by liquid-N2. Another trial was: 2h at -80ºC followed by liquid N2, and the last one was directly to liquid N2. The best results were obtained with the first protocol.

Could anyone give me a hand? Capsules are in good shape and swelling conditions, but the cells are the problem.

Any other conditions I might not be paying enough attention to?

Thanks so much beforehand!!

Ainhoa-
Ainhoa Murua Ugarte
University of the Basque Country
Spain

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Re: Cryopreservation - Encapsulated Cells

Post by x-treme » Sat Jan 14, 2006 11:05 am

Ainhoa M wrote:Hi all!
I am trying several cryopreservation protocols to cryopreserve encapsulated cells (C2C12 mioblasts)but my cell viability results after thawing aren't as good as previous to cryopreservation. In my first trials I used 10%DMSO but after having a 40% decrease in cell viability, I tried higher percentages of DMSO, with even worse results. I am trying 5% and 1% DMSO next week but I am not sure this might be the solution.
I also tried different freezing protocols, such as: 1h at -20ºC, 23h at -80ºC followed by liquid-N2. Another trial was: 2h at -80ºC followed by liquid N2, and the last one was directly to liquid N2. The best results were obtained with the first protocol.

Could anyone give me a hand? Capsules are in good shape and swelling conditions, but the cells are the problem.

Any other conditions I might not be paying enough attention to?

Thanks so much beforehand!!

Ainhoa-



I am not sure - its a trial-u can give0.5% DMSO at the same conditon of the first protocol
or0.5% DMSO at half an hour for -25 degree and 28 hour for -75 degree.
can u mention the temp. of liquid nitrogen u used?

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