The averages of cell replacement by Fah-positive hepatocytes were 2.8% at 6 weeks (6.4% at utmost) and 6.8% at 12 weeks (13.2% at utmost), respectively (Figure 3G). Therefore, human fetal liver cells could xeno-engraft into liver of Fah?/?Rag2?/? mice for liver xeno-repopulation. Figure 3 Human fetal liver cells selleck chem inhibitor have capacity for xeno-engraftment. A: Percentage of E-Cad+ cells in isolated human fetal liver cells by FACS. B: Percentage of E-Cad+ cells after enrichment by FACS; FAH (C; original magnification, ��100) … Livers of primary recipients with high level of repopulation (>10% FAH+ hepatocyte on sections) were perfused to isolate hepatocytes. The level of FAH+ hepatocytes in total isolated hepatocytes was detected by FAH antibody immuno-staining on cytospin slides.

Results indicated that two values about FAH+ hepatocytes were 12.43 �� 2.23% and 11.22 �� 3.12%. Based on this information, we transplanted 3 �� 105 FAH+ hepatocytes into each secondary Fah?/?Rag2?/? recipients. Treatments of anti-asialo GM1 and FK506 were performed as before. At 6 and 12 weeks, liver samples of recipients were harvested and analyzed. The average levels of FAH+ hepatocytes were 8.6% (14.6% at utmost) at 6 weeks and 26.4% (46.7% at utmost) at 12 weeks (Figure 3, E and G). These levels were similar to the levels achieved using adult human hepatocytes. We further confirmed liver xeno-repopulation from human hepatocytes using immunohistochemical analysis with antibody against human albumin.

The levels of human albumin-positive hepatocytes were found to be similar to the levels of FAH+ hepatocytes in both primary (Figure 3, C and D) and secondary Fah?/?Rag2?/? recipients (Figure 3, E and F). Thus the FAH+ hepatocytes were proven to be human hepatocytes. Hepatic Function of Human Hepatocytes after Liver Xeno-Repopulation The recovery of hepatic and metabolic function could be reflected by body weight as a parameter of health status.11 We have found that Fah?/?Rag2?/? recipients with high levels of repopulation gradually gained normal weight by 5 weeks after human hepatocyte transplantation (Figure 4C). By serial staining for hAlb and hAAT with a specific antibody we confirmed the human origin of engrafted hepatocytes and the synthetic function of human hepatocytes (Figure 4, A and B). Figure 4 Engraftment and hepatic function of human hepatocytes after liver xeno-repopulation. Serial staining of human specific Alb Ab (A; original magnification, ��200), and AAT Ab (B; original magnification, ��200) on liver specimen of chimeric … Metabolic parameters were used to analyze the AV-951 recovery of metabolic functions from the replaced human hepatocytes in mouse recipients.