Hi everyone!
For human T cells isolated from PBMCs & frozen after isolation, how would you recommend to proceed with them after thawing? Shall I use them immediately (for sorting for instance) or let them recover (overnight or for a few hours) before experiment?
I need naive T cells, and I m not sure I can maintain this phenotype if I keep them too much in culture.
Thanks in advance for your help :)
Hello Daniela C. Ivan
Once thawed, T cells must be rested for a minimum of 8 hours to a maximum of 16-18 hours to remove any apoptotic cells. A longer duration of resting could separate the dead cells and late apoptotic cells from the lymphocyte population. This would help to increase viability and improve functionality thereby enhancing the specificity and sensitivity of the assay (for instance, flow cytometry assay). The resting process could be helpful to restore the cells as well as have a positive effect on the recovery of the function of lymphocytes.
As you rightly mentioned, resting could also induce phenotypic changes.
Please refer to the research article attached below where in the investigators investigated the influence of resting on the phenotype and functionality of T cells comparing fresh PBMCs as gold standard to rested and non-rested cryopreserved PBMCs.
Article Standardization of cryopreserved peripheral blood mononuclea...
Hope this will be helpful!
Best.
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