Miltenyi, 130-093-631, CD4+CD25+CD45RA+ Regulatory T Cell Isolation Kit, human

Background information
Regulatory CD4+CD25+ T cells isolated from the CD45RA+ naive T cell compartment were shown to be optimal for in vitro expansion. These expanded Treg cells maintained the FoxP3+ phenotype and their suppressive activity, whereas CD45RA– regulatory T cells from the memory T cell compartment lost FoxP3 expression and showed only modest suppressive activity.1

Detailed separation procedure
The isolation is performed in a two-step procedure. First, non-CD4+ and memory T cells are indirectly magnetically labeled with a cocktail of biotin-conjugated antibodies and Anti-Biotin MicroBeads. The labeled cells are subsequently depleted over a MACS® Column. In the second step, the flow-through fraction of pre-enriched CD4+CD45RA+ T cells is labeled with CD25 MicroBeads for subsequent positive selection of CD4+CD25+CD45RA+ T cells.

Downstream applications
CD4+CD25+CD45RA+ regulatory T cells can be isolated from human PBMCs with this kit, for example, for further phenotypical or functional characterization and expansion.

Columns
For the first magnetic separation (depletion): LD or autoMACS® Columns. For the second magnetic separation (positive selection): MS or autoMACS Columns.

Overview
The CD4+CD25+CD45RA+ Regulatory T Cell Isolation Kit was developed for the isolation of CD45RA+ regulatory T cells (Treg) from human PBMCs in a two-step procedure.