Figure 1: Experimental Setup for the Identification of Humoral Transfer Factor(s) From Healthy Volunteers Undergoing RIPC to Isolated Langendorff-Perfused Mouse Hearts

Plasma from 20 volunteers was obtained at 12 pre-defined time points and dialyzed with a dilution of 1:20 for 24 h, using a cutoff of 12 to 14 kDa. After 20 min of stabilization, isolated mouse hearts were perfused with dialysates of human plasma for 15 min, followed by 5 min of wash-out by perfusion with Krebs-Henseleit buffer, before undergoing 20 min of global ischemia. After 120 min of reperfusion, infarct size was determined with 2,3,5-triphenyltetrazolium chloride (TTC) staining, and activation of proteins was analyzed with Western blot analysis. D = dialysate; d = day; I = ischemia; LVP = left ventricular pressure; R = reperfusion; RIPC = remote ischemic pre-conditioning; S = stabilization; WO = wash-out.