Human Regulatory T Cell Expansion And Measurement Of Function

Regulatory T cells (Tregs) are a subpopulation of T cells with the unique ability to suppress the activation of effector T cells and thereby prevent T cell function. The relative frequency of Tregs in peripheral blood is 1–2% of the total lymphocyte population, which is why ex vivo expansion of Tregs prior to clinical applications such as adoptive transfer is necessary. In vitro expansion of Tregs can be triggered by providing the primary and secondary activation signal via crosslinking of the CD3 and CD28 cell surface receptors, and the third activation signal by addition of IL-2 to the culture medium.

This application note describes an optimized protocol for activation and expansion of flow-sorted Tregs with Gibco™ CTS™ Dynabeads™ Treg Xpander. Maintenance of Treg phenotype and function is confirmed by flow cytometric analysis and coculture assay.