Methods in Molecular Biology Volume 821, 2012, pp 279-293
Date: 29 Sep 2011

Expanding Human T Regulatory Cells with the mTOR-Inhibitor Rapamycin

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CD4+CD25+FOXP3+ T regulatory (Treg) cells are pivotal for the induction and maintenance of peripheral tolerance in both mice and humans. The possibility to use Treg cells for the treatment of T-cell-mediated diseases has recently gained increasing momentum. However, given the limited amount of circulating FOXP3+ Treg cells, efficient methods for their ex vivo expansion are highly desirable. Rapamycin allows for in vitro expansion of murine and human FOXP3+ Treg cells, which maintain their regulatory phenotype and suppressive capacity. Here, we describe in detail the powerful methods for enriching human FOXP3+ Treg cells starting from unfractionated CD4+ T cells or for expanding CD25+-enriched Treg cells in the presence of rapamycin.