Hi,
We do transduction on t cells to express a specific target domain. The plasmid expressing the domain has a selecting marker. After selecting the tranduced cells using a specific isolation kit, these cells undergo a REP with irradaited PBMS. I am just missing why and how these feeder cells work to increase the transduced cells?
Thanks
Feeder cells are secreting unknown factors into the media or agar. We still do not know the factors working in these conditions.
Hi
Using allogeneic
PBMCs would provide HLA mismatch environment which adds to the strength of
the stimulatory reaction in addition to their role as feeder cells
Regards
The feeder PBMCs provide factors that help T cell growth, like IL-2, but also, as Zaid said, they induce T cell proliferation due to HLA mismatch. They are irradiated so they will die in the culture after some days, not "contaminating" your culture with unwanted cells after the expansion. To increase the T cell expansion rate and also provide a more homogeneous expansion (not dependent on the HLA mismatch, which will select only a supopulation of your initial T cell repertoire), you can use togeter with the irradiated PBMCS, the Phytohaemagglutinin (PHA). In this way, all your cells should proliferate (MORETTA, A. et al. Direct demonstration of the clonogenic potential of every human peripheral blood T cell. Clonal analysis of HLA-DR expression and cytolytic activity. J Exp Med, v. 157, n. 2, p. 743-54, Feb 1 1983).
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