Cloudz Human Treg Expansion Kit
Cloudz Human Treg Expansion Kit Summary
Includes dissolvable Cloudz CD3/CD28 microspheres that activate and promote the expansion of Treg cells in cell culture.
- Greater than 500-fold expansion of Treg cells from CD4+ T cells in 9 days
- Cloudz CD3/CD28 microspheres are dissolvable
- Easy cell harvesting using concentrated release buffer
- Magnetic bead-free platform
- Optimized for use with R&D Systems cytokines and media
Why Expand Tregs Ex Vivo Using Cloudz Human Treg Expansion Kit?
Forkhead Box P3 (FoxP3)+ regulatory T (Treg) cells are a suppressive subset of CD4+ T cells that function to antagonize immune responses. Treg cells play an important role for preventing autoimmune disease, maintaining immune homeostasis, and modulating immune responses during infection. In vitro differentiation of Treg cells from the larger naïve CD4+ T cell population is often used to obtain large numbers of Tregs for downstream experimentation. Efficient methods of Treg expansion that enable easy cell harvesting are necessary to facilitate basic discovery research and to elucidate their application in preclinical and therapeutic settings.
The Cloudz Human Treg Expansion Kit is designed for the robust expansion of human regulatory T cells (Tregs) cells from CD4+ T cells. The kit contains dissolvable microspheres that are functionalized with anti-CD3 and anti-CD28 antibodies (Cloudz Treg CD3/CD28). The kit also contains Cloudz 6X Release Buffer which quickly dissolve Cloudz Treg CD3/CD28, enabling easy cell harvesting for downstream processing. Each kit contains enough materials to activate and expand FoxP3+ Tregs from 1 x 108 CD4+ T cells, with greater than 500-fold expansion in 9 days.
How to Optimize Treg Expansion using the Cloudz Human Treg Expansion Kit?
This kit was designed as a versatile expansion reagent to support Treg cell expansion in vitro in combination with a variety of media and cytokine combinations. The datasheet contains an optimized protocol for expansion using Recombinant Human IL-2 and ExCellerate Human T Cell Expansion Media. Using the recommended protocol Treg cell can achieve greater than 500-fold expansion in 9 days.
If other cytokines, cytokine concentrations, or media are desired, the expansion protocol may need to be optimized from the provided protocol in the datasheet.
- 16.5 mL of Cloudz Treg CD3/CD28
- 100 mL of Cloudz 6X Release Buffer
Stability and Storage
Upon receipt, this media should be stored at 2-8 °C. Do not use past kit expiration date.
When handling biohazardous materials such as human cells, safe laboratory procedures should be followed and protective clothing should be worn.
- FOR LABORATORY RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES.
- The safety and efficacy of this product in diagnostic or other clinical uses has not been established.
- This reagent should not be used beyond the expiration date indicated on the label.
- Results may vary due to variations among cells derived from different donors.
Fold Expansion of Treg Cells using the Cloudz Human Treg Expansion Kit. Human CD4+ T cells were cultured for 9 days following the protocol and using reagents included in the Cloudz Human Treg Expansion Kit. Expansion of FoxP3-positive Treg cells was evaluated using flow cytometry on Day 5 and Day 9 and fold expansion of was calculated.
Increased FoxP3-positive CD4+ T Cells Following Treg Cell Expansion. Flow cytometry data showing expression of the Treg cell marker, FoxP3, in CD4+T cells on Day 0 (A,C) and Day 9 (C, D) following expansion using the Cloudz Human Treg Expansion Kit. Nine days after expansion, the cells were fixed, permeabilized, and stained with CD4-FITC, CD25-APC, and FoxP3-PE antibodies. Cells in panels (A) and (B) were stained with FoxP3 and CD4. Cells in (C) and (D) were stained with FoxP3 and CD25.
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