Baboon platelets aggregation in different pig cells.

<p>Baboon platelets (2×10⁸) were mixed with (a) pig hepatocytes, (b) pig aortic endothelial cells and (c) pig liver endothelial cells separately (2×10⁶).Mild platelet aggregation was observed, corresponding to 10.8±0.7%, 10.5±0.4%, and 10.4±0.4%. There was no significant difference between these various groups (<i>P</i>>0.05).</p

Pig platelets aggregation in different pig cells.

<p>(a) Pig or (b) baboon platelets (2×10⁸) were stimulated by collagen (0.5 ug/ml) and showed strong aggregation. When pig platelets (2×10⁸) were mixed with (c) pig hepatocytes, (d) pig aortic endothelial cells and (e) pig liver endothelial cells separately (2×10⁶), no platelet aggregation was observed.</p

Cell culture and identification.

<p>Porcine aortic endothelial cells (a), hepatocytes (b), and liver sinusoidal endothelial cells (c) were in culture. The primary cells where then tested by anti- CD31 antibody(d), anti-hepatocyte specific antigen antibody (OCH1E5) (e), and Dil-Ac-LDL up-take(f) and examined under fluorescence microscope.To understand MAC-1 receptor expressed in liver sinusoidal endothelial cells,flow cytometry test for MAC-1 and CD14 in cells was done(g). The expression rate of Mac-1 in cells was 14.2%. In another hand, CD14 expression in cells was negative. It meaned that all the cells were not Kupffer cells.</p

Platelets phagocytosis in different cells.

<p>In (A) CFSE labeled porcine platelets (green) were mixed with pig hepatocytes, aortic endothelial cells and liver endothelial cells (stained by CellTracker™ Blue CMAC, blue), co-cultured for 1 h at 37°C. No phagocytosis was observed (confocal microscopy×400).In (B), CFSE labeled baboon platelets (green) were mixed with pig hepatocytes, aortic endothelial cells and liver endothelial cells (stained by CellTracker™ Blue CMAC, blue), co-cultured for 1 h at 37°C. Platelet internalization by hepatocytes, aortic endothelial cells and liver endothelial cells was absent, mild and strong, respectively (confocal microscopy ×400). In (C), baboon platelets (labeled by CFSE, green) are rapidly internalized by pig liver endothelial cells (stained by CellTracker™7-amino-4-chloromethylcoumarin, blue) (confocal microscopy ×400). The left panel shows the sequential section analysis of a single endothelial cell that has internalized platelets, the sections go from top to bottom. The right panel shows the analysis of one section with simultaneous indication of location of platelets within the endothelial cell, indicating that these are clearly internalized and not on the cell surface.</p