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Human hepatic stem cells transplanted into a fulminant hepatic failure Alb-TRECK/SCID mouse model exhibit liver reconstitution and drug metabolism capabilities.

Zhang RR, Zheng YW, Li B, Tsuchida T, Ueno Y, Nie YZ, Taniguchi H - Stem Cell Res Ther (2015)

Bottom Line: Chimeric rate and survival rate after cell transplantation was evaluated.Expressions of human hepatic-related genes were detected.A human albumin enzyme-linked immunosorbent assay was performed after 50 days of transplantation.

View Article: PubMed Central - PubMed

Affiliation: Department of Regenerative Medicine, Graduate School of Medicine, Yokohama City University, 3-9 Fuku-ura, Kanazawa-ku, Yokohama, Kanagawa, 236-0004, Japan. ranranzhang.0537@hotmail.com.

ABSTRACT

Introduction: Chimeric mice with humanized livers were recently established by transplanting human hepatocytes. This mouse model that is repopulated with functional human hepatocytes could be a useful tool for investigating human hepatic cell biology and drug metabolism and for other preclinical applications. Successfully transplanting human hepatocytes into mice requires that recipient mice with liver failure do not reject these human cells and provide a suitable microenvironment (supportive niche) to promote human donor cell expansion and differentiation. To overcome the limitations of current mouse models, we used Alb-TRECK/SCID mice for in vivo human immature hepatocyte differentiation and humanized liver generation.

Methods: 1.5 μg/kg diphtheria toxin was administrated into 8-week-old Alb-TRECK/SCID mice, and the degree of liver damage was assessed by serum aspartate aminotransferase activity levels. Forty-eight hours later, mice livers were sampled for histological analyses, and the human donor cells were then transplanted into mice livers on the same day. Chimeric rate and survival rate after cell transplantation was evaluated. Expressions of human hepatic-related genes were detected. A human albumin enzyme-linked immunosorbent assay was performed after 50 days of transplantation. On day 60 after transplantation, drug metabolism was examined in mice.

Results: Both human primary fetal liver cells and hepatic stem cells were successfully repopulated in the livers of Alb-TRECK/SCID mice that developed lethal fulminant hepatic failure after administering diphtheria toxin; the repopulation rate in some mice was nearly 100%. Compared with human primary fetal liver cells, human hepatic stem cell transplantation rescued Alb-TRECK/SCID mice with lethal fulminant hepatic failure, and human hepatic stem cell-derived humanized livers secreted more human albumin into mouse sera and also functioned as a "human liver" that could metabolize the drugs ketoprofen and debrisoquine.

Conclusion: Our model of a humanized liver in Alb-TRECK/SCID mice may provide for functional applications such as drug metabolism, drug to drug interactions, and promote other in vivo and in vitro studies.

No MeSH data available.


Related in: MedlinePlus

Extensive mouse hepatocyte proliferation after administration of diphtheria toxin. Immunofluorescent staining for cell proliferation markers Ki67 (A) and BrdU (B) in liver tissues from 8-week-old Alb-TRECK/SCID mice without and with diphtheria toxin (DT) treatment. DT-treated mouse liver with serum aspartate aminotransferase activity of 12,000 IU/L was sampled at 48 hours. At 2 hours before sampling, BrdU (50 mg/kg) was administered intraperitoneally. Experiments were performed with five mice/group, and representative images are shown. Nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI). Scale bars = 100 μm.
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Fig2: Extensive mouse hepatocyte proliferation after administration of diphtheria toxin. Immunofluorescent staining for cell proliferation markers Ki67 (A) and BrdU (B) in liver tissues from 8-week-old Alb-TRECK/SCID mice without and with diphtheria toxin (DT) treatment. DT-treated mouse liver with serum aspartate aminotransferase activity of 12,000 IU/L was sampled at 48 hours. At 2 hours before sampling, BrdU (50 mg/kg) was administered intraperitoneally. Experiments were performed with five mice/group, and representative images are shown. Nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI). Scale bars = 100 μm.

Mentions: To assess in vivo hepatocyte proliferation after the administration of DT, we performed an immunohistochemical analysis using cell cycle markers for total cell cycle activity (Ki-67) and S-phase progression (BrdU incorporation, Cyclin A) in the livers of both normal mice and mice with fulminant hepatic liver failure. This showed that there was a higher degree of Ki67-positive expression (Figure 2A, lower panels) and BrdU incorporation (Figure 2B, lower panels) in livers at 48 hours after the administration of DT. In contrast, no positive Ki67 (Figure 2A, upper panels) and only a few BrdU-positive cells (Figure 2B, upper panels) were detected in normal mouse livers. These results showed that mouse liver regeneration was occurring after DT injection.Figure 2


Human hepatic stem cells transplanted into a fulminant hepatic failure Alb-TRECK/SCID mouse model exhibit liver reconstitution and drug metabolism capabilities.

Zhang RR, Zheng YW, Li B, Tsuchida T, Ueno Y, Nie YZ, Taniguchi H - Stem Cell Res Ther (2015)

Extensive mouse hepatocyte proliferation after administration of diphtheria toxin. Immunofluorescent staining for cell proliferation markers Ki67 (A) and BrdU (B) in liver tissues from 8-week-old Alb-TRECK/SCID mice without and with diphtheria toxin (DT) treatment. DT-treated mouse liver with serum aspartate aminotransferase activity of 12,000 IU/L was sampled at 48 hours. At 2 hours before sampling, BrdU (50 mg/kg) was administered intraperitoneally. Experiments were performed with five mice/group, and representative images are shown. Nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI). Scale bars = 100 μm.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4414454&req=5

Fig2: Extensive mouse hepatocyte proliferation after administration of diphtheria toxin. Immunofluorescent staining for cell proliferation markers Ki67 (A) and BrdU (B) in liver tissues from 8-week-old Alb-TRECK/SCID mice without and with diphtheria toxin (DT) treatment. DT-treated mouse liver with serum aspartate aminotransferase activity of 12,000 IU/L was sampled at 48 hours. At 2 hours before sampling, BrdU (50 mg/kg) was administered intraperitoneally. Experiments were performed with five mice/group, and representative images are shown. Nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI). Scale bars = 100 μm.
Mentions: To assess in vivo hepatocyte proliferation after the administration of DT, we performed an immunohistochemical analysis using cell cycle markers for total cell cycle activity (Ki-67) and S-phase progression (BrdU incorporation, Cyclin A) in the livers of both normal mice and mice with fulminant hepatic liver failure. This showed that there was a higher degree of Ki67-positive expression (Figure 2A, lower panels) and BrdU incorporation (Figure 2B, lower panels) in livers at 48 hours after the administration of DT. In contrast, no positive Ki67 (Figure 2A, upper panels) and only a few BrdU-positive cells (Figure 2B, upper panels) were detected in normal mouse livers. These results showed that mouse liver regeneration was occurring after DT injection.Figure 2

Bottom Line: Chimeric rate and survival rate after cell transplantation was evaluated.Expressions of human hepatic-related genes were detected.A human albumin enzyme-linked immunosorbent assay was performed after 50 days of transplantation.

View Article: PubMed Central - PubMed

Affiliation: Department of Regenerative Medicine, Graduate School of Medicine, Yokohama City University, 3-9 Fuku-ura, Kanazawa-ku, Yokohama, Kanagawa, 236-0004, Japan. ranranzhang.0537@hotmail.com.

ABSTRACT

Introduction: Chimeric mice with humanized livers were recently established by transplanting human hepatocytes. This mouse model that is repopulated with functional human hepatocytes could be a useful tool for investigating human hepatic cell biology and drug metabolism and for other preclinical applications. Successfully transplanting human hepatocytes into mice requires that recipient mice with liver failure do not reject these human cells and provide a suitable microenvironment (supportive niche) to promote human donor cell expansion and differentiation. To overcome the limitations of current mouse models, we used Alb-TRECK/SCID mice for in vivo human immature hepatocyte differentiation and humanized liver generation.

Methods: 1.5 μg/kg diphtheria toxin was administrated into 8-week-old Alb-TRECK/SCID mice, and the degree of liver damage was assessed by serum aspartate aminotransferase activity levels. Forty-eight hours later, mice livers were sampled for histological analyses, and the human donor cells were then transplanted into mice livers on the same day. Chimeric rate and survival rate after cell transplantation was evaluated. Expressions of human hepatic-related genes were detected. A human albumin enzyme-linked immunosorbent assay was performed after 50 days of transplantation. On day 60 after transplantation, drug metabolism was examined in mice.

Results: Both human primary fetal liver cells and hepatic stem cells were successfully repopulated in the livers of Alb-TRECK/SCID mice that developed lethal fulminant hepatic failure after administering diphtheria toxin; the repopulation rate in some mice was nearly 100%. Compared with human primary fetal liver cells, human hepatic stem cell transplantation rescued Alb-TRECK/SCID mice with lethal fulminant hepatic failure, and human hepatic stem cell-derived humanized livers secreted more human albumin into mouse sera and also functioned as a "human liver" that could metabolize the drugs ketoprofen and debrisoquine.

Conclusion: Our model of a humanized liver in Alb-TRECK/SCID mice may provide for functional applications such as drug metabolism, drug to drug interactions, and promote other in vivo and in vitro studies.

No MeSH data available.


Related in: MedlinePlus