I need to transport PBMCs which I had frozen in liquid nitrogen to another Lab where I will carry out flow cytometry analysis of some cells, as well as Elispot for T cell secretion of IFN-Y.
Due to the cost of transporting these cells by air in a liquid nitrogen container, I am wondering if transporting the cells in dry ice will affect the assay I want to carry out. If so, can someone suggest any alternative to shipping frozen PBMC in liquid nitrogen.
Thank you
Dear Ihedinachi,
The cell transporting in dry ice should not be a problem for those functional assays, specially if your PBMC samples have a considerable frequency of antigen-specific T cells. Even with dead cells after thawing cells, you would still be able to detect functional T cells in this case. However, if the T cell frequency is too low, you may have to expand them before running those assays.
Eight years ago, I had to culture frozen PBMCs from rhesus macaques with IL-2 and IL-7 before the functional assays. That strategy helped me to expand functional rare T cell clones that I detected through ELISPOT and FACS.
Below you can find the paper with all culture info:
- SILVEIRA ELV, Fogg MH, Leskowitz RM, Ertl HC, Wiseman RW, O'Connor DH, Lieberman P, Wang F, Villinger F. Therapeutic vaccination against the rhesus lymphocryptovirus EBNA-1 homologue, rhEBNA-1, elicits T cell responses to novel epitopes in rhesus macaques. Journal of Virology, v. 87, p. 13904-13910, 2013.
Best of luck,
Eduardo L. V. Silveira
If you need high vitality and full preservation of T cell function, I do not think there are many alternatives to liquid nitrogen.. dry ice will do if the time of transportation is relatively short.
I suggest you carry out a test in your lab with an expendable sample by simulating your transport method (ie. take the vial, place in the same container and dry ice amount you would use for your final experiment and keep them for the expected time of travel at the expected temperature). After the "simulated" travel, thaw your cells as you would do for the real experiment and check vitality and recovery number (if you can, use the same read out you will use with the real samples).
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