Hello everybody,
We would like to address the function of our gdT cells in vitro.
These cells are isolated by positive selection from murine lungs and we would like to assess there antitumoral activity in vitro against cancer cell line.
We culture our magnetic-sorted gd T cells 24 hours in either in:
- DMEM FCS10% L glu PS NaPyr NaBicarbonate
or
- RPMI FCS10% PS L Glu beta-mercapto 50uM
and our viability rate goes down from 95% to less then 15%.
Any advice?
Thank in advance.
Hello Alexis Broquet
You could use cytokine-based approach for expansion of γδ-T cells in culture. Please refer to the articles I have attached below for your reference.
https://www.sciencedirect.com/science/article/abs/pii/S0022175922001090?via%3Dihub
https://www.immunologyresearchjournal.com/articles/expansion-of-gamma-delta-t-cells--a-short-review-on-bisphosphonate-and-k562based-methods.pdf
Best.
Thanks Malcolm for your fast reply.
Is there a way to keep gd T cells alive without the expansion phase described in these papers.
I will already have enough cells to test their function so I won't need that expansion phase before incubating them with cancer cells.
Hello Alexis
You may try low dose of IL-2 say (100 U/ml). It may help to keep them alive for a few days hopefully.
ok. thanks. I'll try as just a regular seems to be not suffisent to maintain acceptable viability rate.
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