Recently my group become interested in investigate the role of Innate cells ILCs and NKT in human asthma e we are now collecting data for a study in asthma severity, including the collection of blood and sputum for measuring cytokines in PBMC cell supernatant. However we do not have reagents for studding cell of innate immunity by cytometry and it will takes months in Brazil to obtain such reagents. Then I would like to have a good protocol to cryopreserve blood or PBMC and sputum cells for assay them in a few month for markers of ILCs and NKT cells. Then my query is what is the best way to preserve these cells for this purpose?
Thanks,
Hi Neuza, the standard way to cryopreserve cells is to incubate them with 10% DMSO in serum and freeze them slowly in -80C freezer. (basically, wrap vials containing cells resuspended in 10% DMSO, with lots of tissue paper and put the vials into a -80C. freezer.
The frozen cells will keep for months at -80C.
Thanks a lot Steingrimur. Do you mean that you have assayed frozen ILCs and NKT cell by cytometry using this protocol and it worked well?. I need to be sure in order to order the reagents.
Best regards,
Neuza
Hi Neuza, sorry, I have never assayed frozen ILCs and NKT cells and analyzed them by cytometry.
I have frozen whole PBMCs layers isolated from buffy coats using 10% DMSO and measured intracellular kinase levels from B-cells using ELISA, even after months of storage at -80C.
Sorry I can't be of more help.
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