Determine the fate of your T cells

Human T cells were isolated from peripheral blood mononuclear cells (PBMCs) using the Pan T Cell Isolation Kit, human, and stimulated with the T Cell Activation/Expansion Kit, human. Cells were cultivated in the presence of IL-2 for 14 days in either TexMACS GMP Medium, TexMACS Medium, research grade, a competitor’s T cell medium, or basal medium (RPMI + 10% serum). CD4 and CD8 expression was analyzed via flow cytometry on a MACSQuant® Analyzer and cell expansion was evaluated by cell counting at culture onset and after 14 days of culture. Highest expansion rates were achieved when cultivating T cells in TexMACS Medium (MACS GMP and research grade) compared to a competitor’s T cell medium or basal medium.

Human and mouse T cells were isolated using the respective Pan T Cell Isolation Kits. Purified T cells were activated with the respective T Cell Activation/Expansion Kit and cultured in basal medium (RPMI + 10% serum) in the presence of IL-2 for the indicated time periods. CD4 and CD8 expression was analyzed via flow cytometry on a MACSQuant® Analyzer and cell expansion was evaluated by cell counting at culture onset and after indicated days in culture.

Human T cells were isolated using the Pan T Cell Isolation Kit, human. Purified T cells were stimulated with T Cell TransAct, human and cultured in TexMACS Medium containing IL-2 for the indicated time period.  CD4, CD8, CD25, and CD69 expression was analyzed via flow cytometry on a MACSQuant® Analyzer. Proliferation was measured using the cell division tracker CFSE.  T cells activated with T Cell TransAct, human showed high expression of activation markers CD25 and CD69 after 48 h, as well as strong proliferation after 7 days of culture.

Activity of Human IL-2 IS, premium grade, (red bar) was compared to another commercially available product (black bar).

Naïve mouse CD4⁺ T cells were isolated using the Naïve CD4⁺ T Cell Isolation Kit, mouse, activated via the T Cell Activation/Expansion Kit, mouse and polarized into T_H1, T_H2 and T_H17 lineages using the respective CytoBox as indicated. Cells were cultured either in TexMACS Medium (+ 10% serum) or RPMI (+ 10% serum) for 5 days . Analysis of lineage-specific transcription factors was performed via flow cytometry on a MACSQuant® Analyzer.  In line with the expected characteristic cytokine expression profiles, the T_H1 culture showed a high percentage of IFN-γ–producing, but no IL-4– producing cells. The T_H2 culture featured a high frequency of IL-4–, but no IFN-γ–expressing cells, whereas the T_H17 culture was characterized by a high level of IL-17–producing cells and the absence of IFN-γ–producing cells. Furthermore, in vitro mouse T_H cell subset differentiation in the presence of TexMACS Medium led to higher expression levels of characteristic effector cytokines than culture in RPMI medium.

Mouse T_H cell subsets and corresponding polarizing cytokines, surface markers, transcription factors, and secreted cytokines.