The increasing interest in the tumor microenvironment leads to a focus on new bioassays to represent all the players of the cancer immune response. Some of these players like regulatory T cells play an important role by downregulating the anti-tumor response. Their regulation mechanisms constitute an important target for new therapeutics. In order to study these mechanisms in a human model, suppressive Treg bioassays mimicking the suppressive action of these cells were developed and optimized. In vitro suppressive assays come with many technical challenges, therefore protocols for the purification and in vitro culture of regulatory T cells were optimized and fine-tuned to result in an optimal assay window e and allow screening of multiple candidates. Donor-to-donor variation is controlled by pre-evaluation of multiple donors and standardized methods are used for isolation and in vitro culture of responding and suppressive cells. Access to a large and broad panel of healthy donors is required for the evaluation of therapeutic agents targeting the regulatory T cell pathway.
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