Hi everyone,
I am currently working on the analysis of T lymphocyte sub populations in breast cancer tissues using flow cytometry, currently I have frozen breast tissues in DMSO 10 % SVF, and i'm wondering what could be the best defrosting protocol to follow without losing many cells..?
Thanks in advance
Dear Dounia,
I totally agree with Marica.
You should use the classic protocol of rapid thawing at 37°C water bath.
It is important for cell viability that the cells are thawed and processed quickly; thawing should take only a minute or two.
After that add enough quantity of pre-warmed growth medium (it is crucial to be pre-warmed at 37 C before the addition).
Transfer the tissue to a 50 ml conical polypropylene tube and add additional quantity of chosen growth medium.
Then you will be ready for the next procedure (seeding).
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