Hello every one,
I want to conduct an adoptive transfer experiment to check the CD8 T cell populations in B6 mice from OT-1 mice. What should be the ideal number of cells required for the transfer (for an i.v injection)
Thank you.
It depends on what exactly you would like to do. For doing in vivo T cell proliferation, we normally inject 10-50 million cells per animal in 200 microlitres of PBS. With this number you can clearly see them homing and proliferating in lymph nodes and spleen. Follow the attached article for more information. Good luck
The adoptive transfer of T cells has been a matter of debate for as long as T cells are transferred and accordingly, there are many different protocolls used in the scientific comunity.
While we used to inject about 1x106 T cells (OTI) per mouse (in 200ul PBS), other groups (e.g. the group of Ton Schumacher) successfully transferred 1000 OTI cells.
Even more fascinating was the finding by the group of Dirk Busch in Munich, who could show that already the injection of ONE T cell results in a T cell pool that is not different from the pool generated after injection of many more T cells.
Stemberger C, Huster H, Koffler M, Anderl F, Schiemann M, Wagner H, Busch DH: “A single naive CD8+ T cell precursor can develop into diverse effector and memory subsets”. Immunity. 2007; 27(6): 985-997
So, injecting fewer T cells saves lifes ;-)
I agree in both the answers above. In our lab transfer of 5-7 million cells/100ul has worked well for us. It depends upon the objective of experiment and to some extent technical expertise.
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