Figure 10

hMSCs from three different donors effect T cell activation and cytokine production. Murine CD8⁺ T cells (8.3 TCR transgenic T cells) were isolated and purified from lymph nodes and spleens of NOD 8.3 TCR transgenic mice using Miltenyi mouse CD8⁺ T cell isolation kit and cultured for three days in the presence of specific peptides and antigen-presenting cells (APCs). Human bone marrow-derived mesenchymal stem cells (hMSCs) from three different donors (donor A, B and C) were added to the wells at the beginning of the culture at a ratio of 5:1 (T cells:hMSC). The cells were harvested, activation markers (CD25, CD69) were evaluated by flow cytometry and the percent inhibition of activation-induced mean fluorescence intensity (MFI) increase was calculated as follows: (MFI increase of activated T cells without MSCs - MFI increase of T cells with MSCs)/MFI increase of activated T cells without MSCs × 100%. Percent of inhibition was shown for all three donors (panel A). Cytokine concentrations in the culture supernatant were analyzed using a multiplex ELISA assay and the percent inhibition in cytokine production was shown for all three donors (panel B). The graph is representative of two independent experiments with similar results.