Pure and Functional Human Tregs
- Isolate pure CD4+CD25+ cells with high expression of Foxp3
- No antibody bound to CD25 after cell isolation
- Simple cell isolation, no columns
The new Dynabeads® Regulatory CD4+CD25+ T Cell Kit is an easy-to-use, flow-compatible kit for positive isolation of human regulatory T cells (Treg cells). This tube-based technology gives you highly functional Treg cells with up to 97% purity (CD4+CD25+ expression); on average over 80% of the isolated CD25+ cells express the transcription factor Foxp3.
With this simple 3 step magnetic isolation procedure (figure 1) cells are left both bead- and antibody-free after isolation, and can be used in any cell-based downstream experiment. Due to the Dynabeads polymer coating and the fact that the beads are completely removed after isolation, your cells are exposed to minimal stress and there will be no foreign structures from the isolation method left with your cells after isolation.
Regulatory CD4+CD25+ T cells are a specialized subpopulation of T cells that act to maintain homeostasis within the immune system. Recent advances in the characterization of this cell population have firmly established their existence and their critical role in keeping the balance of the immune response. Evidence from experimental mouse and human models have demonstrated that the immunosuppressive potential of these cells can be utilized in the treatment of various diseases such as autoimmunity, infectious diseases and cancer.
A. In the PBMC sample, 2.5% of the cells were CD4+CD25+. B. After negative isolation of CD4+ cells, 7% of the CD4+ cells expressed CD25. C. After isolation of CD25+ cells using Dynabeads CD25, 1% of the low expressing CD25+ cells remained in the CD4+CD25- fraction. D. The isolated CD4+CD25+ cells were 97% pure.
Tube-based vs. Column-based Treg isolation
Higher number of Foxp3+ cells using Dynabeads.Treg cells were isolated from two different donors using Dynabeads Regulatory CD4+CD25+ T Cell Kit (left) or a column-based method (right). Treg purity was identified by expression of CD25 and Foxp3 after isolation.
For Specific Results of:
- Suppressive capacity of isolated Tregs
- Expansion of isolated Tregs
- Suppressive capacity of expanded Tregs
Figure 1. The 3 step procedure for isolation of Tregs from PBMC with Dynabeads Regulatory CD4+CD25+ T Cell Kit. First, untouched CD4+ T cells are negatively isolated using Ab mix and Depletion Dynabeads (step 1). Next CD25+ cells are positively isolated using Dynabeads CD25 (step 2), and cells are released from beads using DETACHaBEAD® (step 3).
Treg isolation & expansion (poster)