Figure 1: Experimental design of study. Five HLA-class I mismatched BMMSC and ASC pairs were used. BM-MSC and ASC were either unstimulated or pre-stimulated for 1 week with IFNγ. To obtain educated CD8+ T cells, a coculture was established with BM-MSC or ASC and HLA-class I mismatched PBMC in the presence of 200 U/ml IL-2. CD3+CD8+ effectors (BM-MSC educated CD8+ T cells; IFNγ-BM-MSC educated CD8+ T cells; ASC educated CD8+ T cells and IFNγ-ASC educated CD8+ T cells) were selected via FACS sorting. CD3+CD8- cells were isolated from all co-cultures as a negative control. After Europium labeling, target cells, either IFNγ-stimulated or unstimulated BM-MSC and ASC, were exposed to the effector cells as depicted here for BM-MSC educated CD8+ T cells. Europium release was assessed as a measure of CD8+ T cell mediated lysis.