hi all,
if I preserve monocyte in DMSO and FBS today. can I use one of them the next day and out them in culture to do an experiment.
Historically, it has been stated that one should wait 1 to 2 weeks after freezing to use cells. However, we have all used them a day or two later with some success. If this study is important I would suggest that you qualify the cells/timing by comparing the outcome parameters 24 hrs and 14 days post freezing. Of course it is best if you use a critical point freezer or a Mr Frosty to optimize the freezing process.
On a different note, if you are going to use cells the next day store them at 4C over night after diluting 1:4 in complete media. Easier and better approach.
In my experience, neither approach is ideal. I would recommend setting up the assays the same day regardless of how long the whole assay setup takes. Monocytes do not freeze well and you may have trouble reviving sufficient number of cells for your assay. Leaving them overnight at 4C is also not a good idea as monocytes do not survive for long post-isolation, unless you culture them in the presence of stimulants. They also tend to stick to surfaces which would make recovery difficult.
Please see the following Mx by McMannis. We also qualified both frozen cells and cells from an apheresis that were shipped overnight for monocyte isolation with success. Lemarie, C. et al., 2007. Purification of monocytes from cryopreserved mobilized apheresis products by elutriation with the Elutra device. Journal of immunological methods, 318(1-2), pp.30–6.
I apologize if I understood it incorrectly. I agree that monocytes can be isolated easily from both fresh and frozen apheresis/whole blood samples. I thought Imene Jridi wanted to freeze isolated monocytes, thaw them and then put those frozen-thawed monocytes in culture. In my experience, once you start isolating rare cell subsets (monocytes, Treg and so on), freezing them for future use is not a good idea.
We don't normally freeze monocytes but often freeze apheresis products. However, we do freeze transduced DCs for use as a clinical vaccine and TILs with success.
Gulen, D. et al., 2008. Closing the manufacturing process of dendritic cell vaccines transduced with adenovirus vectors. International immunopharmacology, 8(13-14), pp.1728–36.
Gülen, D. et al., 2012. Cryopreservation of adenovirus-transfected dendritic cells (DCs) for clinical use. International immunopharmacology, 13(1), pp.61–8.
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