Dear All,
Will cryopreservation affect TCR sequence?
My plan is to store our PBMC at -80 degree first, and these cells will later be thawed and sorted by FACS for TCR sequencing.
Thank you,
Hello
See link here, Hope help you
Article Homeostasis of lymphocytes and monocytes in frequent blood donors
Hi,
In our work, the monocytes could be frozen in this way for about 3 months (we have not used longer periods, but they are possibly possible)
Freezing was performed immediately after isolation (ficall/microbead CD14+). When defrosting, make sure to transfer to medium as quickly as possible and perform a washing step.
wir benutzen RPMI-1640 Medium mit 10% FCS, 2mM L-Glutamin und Pen/Strep.
Einfriermedium:
30ml RPMI Medium
10ml DMSO
10ml FCS
good luck
As you plan on sequencing the TCR, vital freezing of PBMCs is not expected to impact your analysis (as you will be working on genomic DNA). The only bias you should take into account is the risk of altering the relative abundance of the subpopulations you will be extracting DNA from (but this is more likely affected by FACS sorting than by the freeze-thaw procedure).
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