CellXVivo Mouse Th17 Cell Differentiation Kit

For the differentiation of Th17 cells from a preparation of CD4+ T cells isolated from mouse splenocytes.

Why Expand Th17 Cells Ex Vivo?

T helper type 17 (Th17) cells are a subset of CD4⁺ effector T cells that promote cell-mediated immune responses against extracellular bacteria and fungi. Differentiation into the Th17 lineage is promoted by cytokines such as TGF-beta and IL-6, while their survival and expansion are dependent on IL-21 and IL-23. Th17 cells secrete TNF-alpha, IL-6, IL-9, IL-17A, IL-17F, IL-21, and IL-22. Th17 polarized cells are present in low abundance in normal mouse spleen and peripheral blood. In vitro differentiation of mouse Th17 cells from the larger naïve CD4⁺ T cell population provides increased numbers of Th17 cells to facilitate downstream research.

This kit contains the following reagents for the ex vivo differentiation of mouse Th17 cells.

• Hamster Anti-Mouse CD3
• Rat Anti-Mouse CD28
• Mouse Th17 Reagent 1
• Mouse Th17 Reagent 2
• Mouse Th17 Reagent 3
• Mouse Th17 Reagent 4
• Mouse Th17 Reagent 5
• Reconstitution Buffer 1
• Reconstitution Buffer 2
• Reconstitution Buffer 3
• 20X Wash Buffer

The quantity of the components in the kit is sufficient to differentiate approximately 50x10⁶ naïve CD4⁺ T cells, and generate ~200 x 10⁶ cells, of which >40% are Th17 polarized cells within your starting CD4⁺ T cell population.

Note: Results may vary due to strain age and/or the health of the mice used for isolation.

Stability and Storage

Store the unopened kit at ≤-20 °C. Do not use past the kit expiration date. *Provided this is within the expiration date of the kit.

Limitations

• 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.

T helper type 17 (Th17) cells are involved in the immune response mounted against specific fungi and extracellular bacteria. In mice, Th17 cells develop from naive CD4⁺ T cells in the presence of TGF-beta and IL-6. These cytokines induce the STAT3-dependent expression of IL-21, IL-23 R, and the transcription factor, ROR gamma t. IL-21 and IL-23 regulate the establishment and clonal expansion of Th17 cells, while ROR gamma t-induced gene expression leads to the secretion of IL-17A, IL-17F, and IL-22. Cytokines secreted by Th17 cells stimulate chemokine secretion by resident cells, leading to the recruitment of neutrophils and macrophages to sites of inflammation. These cells, in turn, produce additional cytokines and proteases that further exacerbate the immune response. In contrast to mouse Th17 differentiation, Th17 polarization in humans requires IL-1 beta, IL-6, IL-21, and IL-23, but seems to be less dependent upon TGF-beta. One other notable difference is that human Th17 cells secrete IL-26, an IL-10 family cytokine without a murine homologue. Cytokines produced by Th17 cells can have both beneficial and pathogenic effects. While they play a central role in eliminating harmful microbes, persistent secretion of Th17 cytokines promotes chronic inflammation and may be involved in the pathogenesis of inflammatory and autoimmune diseases, including rheumatoid arthritis, multiple sclerosis, and inflammatory bowel disorders.