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. 2018 Sep 6;8:13351. doi: 10.1038/s41598-018-31569-1

Figure 2.

Figure 2

Mixed lymphocyte reactions and flow cytometry analysis of CD25 expression on lymph node cells (LNCs) in response to allogeneic donor cardiosphere-derived cells (CDCs). Initial titration experiments revealed a peak stimulation of LNCs at between 1:100 and 1:1000 ratios of CDCs:LNCs (n = 3, with triplicate samples in each a). Different donor CDCs cultured with LNCs, at a ratio of 1:500 CDCs:LNCs. At this ratio, there was no difference between resting LNC proliferation and those cultured with allogeneic CDCs (n = 4 dogs, with triplicate samples for each, p > 0.05 b–g). FACS analysis of co-cultured cells allowed LNCs to be gated h. Graphs i–k show only gated LNCs with simultaneous CD25 and PI staining analysed for both LNC activation and death respectively. ConA treated LNCs had more CD25 signal compared to LNCs alone without stimulation (n = 4, p < 0.001, i and j). Co-culture LNCs with CDCs had less CD25 levels than LNCs alone or LNCs stimulated with ConA (n = 3, p < 0.001, k). Bars represent mean ± SEM of 3 different dogs for a, and mean ± SEM of triplicate results in each b–g.