As far as I know, there are too many methods to freeze stem cell lines and to inhibit cell death during frozen time in long intervals, some researches reported that using specific cell death inhibitors (e.g. caspase-1 inhibitor, zVAD-fmk) can reduce potential cell death occurring in long freezing times. Furthermore, we also know that during thawing process, activation of caspases can induce apoptosis contributing to cryo-injury to transplant grafts. Is there any suggestion to prepare better freezing media for HPSCs? Which media can be better specifically for HPCs?
Don't have any personal experience with freezing HSCs but seems there are a few specialized freezing media out there that might work.
http://www.amsbio.com/cellbanker-freezing-media.aspx (HSC-Banker)
https://www.stemcell.com/products/product-types/cell-storage-media.html (BloodStor 55-5)
https://www.miltenyibiotec.com/CA-en/search.html?search=cryo-brew (HSC-brew)
Thank you, I am mostly searching for alternative known ingrediented prepared versions. I already searched for those freezing media that you sent.
We have simply used 5% DMSO and 95% FBS. This seemed to work better than any commercial freezing media we have tried. We do see cell death but we get about 70% recovery from the amount of cells we freeze. The cells we recover upon thaw give viability in the 90-96% range and work well in various experiments looking at protein function and message. We also do the dropwise thawing method.
90-95% FBS and 10-5% DMSO should work well otherwise a more expensive commercially available alternative like StemMACS™ Cryo-Brew, 130-109-558, Miltenyi Biotech, could work.
In my experience with umbilical cord blood we use DMSO Dextran commercial medium, but we are studding a mixture of DMSO Dextrose as a suitable alternative. In our results we recovered about 80-90% of CD34+ viable cells thawing just with PBS or saline. Our results are in revision for publication at this moment, I hope they will be available soon.
Let me know if you need more info
Diego
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