Immunological Compatibility of Bone Tissues from Alpha-1,3-galactosyltransferase Knockout Pig for Xenotransplantation

Figure 5

Effect of bone extracts on the percentage of CD4⁺ T cells and regulatory T cells. PBMCs were obtained from heparinized blood of healthy volunteers by histopaque gradient centrifugation. Bone extracts were added to PBMC cultures at 100 ug/ml concentration after 96 hrs, cells were stained for CD3 and CD4 using specific antibodies, and percentage of CD4⁺ T cells was assessed by flow cytometry analysis (a). Cells were further stained for intracellular expression of foxp3 and percentages of Tregs in CD4⁺ T cells were analyzed using flow cytometry analysis.. #, ##, and ### indicate significant difference from the control group. p < 0.05, p < 0.005, and p < 0.001 indicate significant difference from the WT group. Experiments were repeated at least three times independently with hPBMC from 3-4 blood donors and bone extracts from 2-3 pigs. Data are expressed as means ± SEM.

(a)

(b)