I'm having some problems with cryopreservation of BY-2 suspension cells. I followed the protocol from "Initiation, growth and cryopreservation of plant cell suspension cultures, Mustafa et al. (2011)" but I only get dead cells after thawing (in attachment I added some pictures of the cells with and without Evans blue staining).
I also halved the concentration of mannitol in the pre-culture medium because the cells where stressed with the mannitol concentration in the paper. However, after thawing the cells again looked unhealthy and I got no regrowth in the medium after more than 1 week.
Thank you very much in advance.
Kind regards, Jonas
I'm not sure if helping this advice:
try to use DNAse after thawing (as we used for frozen blood cells).
Before use ice cryoprotector like DMSO and freeze one degree per minute
García-Piñeres AJ, Hildesheim A, Williams M, Trivett M, Strobl S, Pinto LA. DNAse treatment following thawing of cryopreserved PBMC is a procedure suitable for lymphocyte functional studies. J Immunol Methods. 2006 Jun 30;313(1-2):209-13. Epub 2006 May 17. PubMed PMID: 16737707.
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