We have recently utilised Bambanker (BB01) from Nippon Genetics (Geneflow in UK), which is a serum free DMSO containing cryogenic storage medium for cells. PBMCs (5x10e5/ml) were not happy in this medium when stored at -80C (in an Isopropanol container, althouth I know this is not necessary) for ~12 months (viability dropped considerably from baseline and 1 month data, measured using flow cytometry) and thus we have moved them to Liquid N2. Since then I have found out that Nippon Genetics do not have data on PBMCs rather immortalised cell lines. Moreover I am still not sure whether they use viability and/or growth rate to validate its use.
I am writing to find out other peoples experiences of Bambanker. Going forward I plan to place PBMCs into Bambanker, freeze down at-80 then transfer to Liq N2. Does anybody else do this and have they experienced good (>90% viability?). Thanks Adam Wright
I never have used that cryopreservative, but it certainly is no surprise that survival was low after 12 months at -80C. Long term survival storage of eukaryotic cells requires temperatures lower than -80C. Your plan for initial freezing in a -80C freezer followed by transfer to LiN is the normal strategy and should provide much better long term survival.
Mammalian cell do fine in Bambanker at -80°C. We've used it for years and have had no problems. PBMCs don't do as well in this freezing medium, however.
Michael Campa For what duration of time were you able to recover cells from -80C?
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