Determine the fate of your T cells

This complete workflow allows for a reliable and efficient T helper cell (TH cell) differentiation. Starting with cell isolation of naive CD4+ T cells over to in vitro activation and differentiation, up to multiparameter flow cell analysis.

We provide gentle cell separation strategies, as well as reliable cell culture reagents such as T cell activation tools, T cell media and polarizing cytokines for the efficient differentiation of TH1, TH2, and TH17 cells.

Isolation of naïve CD4+ T cells

Choose from a variety of MicroBeads and T cell Isolation kits for the separation of human and mouse naïve CD4+ T helper cells.

Naive T Cell Isolation Kits mouse and human

The Naïve T Cell Isolation Kits were developed for the isolation of untouched naïve T cells from human peripheral blood or mouse spleen single cell suspensions. Naïve T cells are ready for downstream applications, e.g., in studies on naive T helper cell activation and differentiation as well as and in functional studies of DCs.

Isolation of untouched naive CD4+ T helper cells from PBMCs using the Naive CD4+ T Cell Isolation Kit II, human. Cells were stained with CD4 and CD45RA antibodies and analyzed by flow cytometry using the MACSQuant® Analyzer. 

Naïve CD4+ T cells were isolated from a single-cell suspension from mouse spleen using the Naive CD4+ T Cell Isolation Kit, mouse. Cells were fluorescently stained with CD45-VioGreen, CD4-VioBlue®, CD62L-PE, CD3ε-APC-Vio770™ , and CD44-FITC and analyzed by flow cytometry using the MACSQuant® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence and a gate was set on CD45+ CD3e+ CD4+ cells. 

CD62L, CD45RA and CD45RO MicroBeads 

Combine pre-enrichment with the CD4+ T cell Isolation Kits by positive selection or depletion with the respective MicroBeads to isolate even the rarest naïve CD4+ T cell populations.

Scientific poster
T helper cells: a complete workflow for cell preparation, isolation, stimulation, polarization and analysis

Christian Neumann1,2, Alexander Scheffold2, Nicole Pietz3, Anna Foerster-Marniok3, Michaela Niemöller3, Julia Milleck3, Anne Richter3, and Tobias Schenk
1German Rheumatism Research Centre Berlin, an Institute of the Leibniz-Association, Berlin, Germany; 2Department of Rheumatology and Clinical Immunology, Charité University Hospital Berlin, Germany; 3Miltenyi Biotec B.V. & Co. KG, Bergisch Gladbach, Germany

T cell media

TexMACS™ Medium is an optimized, serum-free cell culture medium, developed for reliable T cell and Treg cell culture results. Chemically-defined and without animal-derived components, it allows for the highest reproducibility in human and mouse cell culture applications.

Comparison of expansion rates of human CD4+ and CD8+ T cells in TexMACS Medium or competitor's T cell medium using the T Cell Activation/Expansion Kit, human

TexMACS Medium

  • Free of serum and other animal-derived components                                                         
  • Reliable T cell activation and high expansion rates
  • Up to MACS® GMP grade
  • Available with or without phenol red
     

T cell activation

We offer a variety of polyclonal T cell stimulation tools for you to choose from. Find the options below:

Expansion of human and mouse CD4+ and CD8+ T cells using T Cell Activation/Expansion Kits

T cell Activation/Expansion Kits 

  • MACSiBead™ Particles loaded with functional-grade antibodies
  • Efficient activation and expansion of human, mouse, and non-human primate T cells or Treg cells.
Expansion of human CD4+ and CD8+ T cells with T Cell TransAct

T Cell TransAct™ Stimulation Reagent

  • Ready-to-use reagent for activation and expansion of human T cells                                      
  • Simple volumetric dosage and easy removal by washing
  • Up to MACS GMP grade

We provide a broad selection of superior-quality cytokines, with high biological activity at a competitive price. Three different grades, research, premium, and MACS® GMP grade, provide flexibility in your assay setup.

Human IL-2 IS biological activity

MACS® Premium-Grade Cytokines 

  • Well-defined biological activities, normalized to international reference standards (IU/mg)
  • Exact unit dosing for reproducible results
  • Lot specific activity, requiring no laborious pre-testing
Mouse TH cell polarization using CytoBoxes (TH1, TH2, or TH17) in combination with either RPMI or TexMACS Medium

TH cell polarization with CytoBoxes 

  • Convenient all-in-one starter kits for polarization of mouse TH1, TH2 and TH17 cells
  • Recombinant cytokines and functional-grade blocking antibodies
Principle of the Cytokine Secretion Assays

Detection and enrichment of cytokine-producing T cells

Cytokine secretion assays are designed for highly sensitive detection and magnetic enrichment of viable cytokine-secreting cells. This unique technology enables detection of one antigen-specific T cell per million cells. Isolated cells can be immediately used for downstream applications, e.g., cell culture and flow cytometric analysis.

A range of kits are available for human and mouse cells secreting TNF-α, GMCSF, IFN-α, IFN-ƴ, or IL-2, -4, -5, -10, -12, -13, -17, -22.

Cytokine Secretion Assay –  Detection Kits 

  • Accurate analysis of cytokine secretion at single-cell level
  • Sensitive detection of one T cell per million cells

Cytokine Secretion Assay – Cell Enrichment and Detection Kits

  • Unique technology for the isolation of viable cytokine-secreting cells
  • Additional magnetic enrichment step
Mouse TH cell subsets and corresponding polarizing cytokines, surface markers, transcription factors, and secreted cytokines.
Human TH cell subsets and corresponding polarizing cytokines, surface markers, transcription factors, and secreted cytokines.

Multiparameter flow analysis of T cells

As the final step in most workflows, flow analysis plays an important part in various T cell applications. Our flow reagents can help you ensure that the data you generate on your flow cytometer is reliable and consistent:

  • REAfinity™ Recombinant Antibodies – recombinantly generated antibodies that ensure lot-to-lot consistency and eliminate background signal
  • MACSplex™ Cytokine Kits – analysis of multiple soluble cytokines in one sample using any standard flow cytometer
  • Rapid Cytokine Inspector – combined surface marker and intracellular cytokine staining for multiparameter analysis of activated human T cells
  • Cytokine Secretion Assay – Detection Kits – flow analysis of cytokine secretion at a detection level of one T cell in a million

Build your own T cell flow panel with our Flow Panel Builder

Application note
Flow cytometry analysis of TH subsets

Find out more about multiparameter flow analysis of your favorite TH cell subsets.
 

Get acquainted with our protocols and antibody panels for your workflow in T cell research.

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