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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

Alb-TRECK/SCID mice develop fulminant hepatic failure after diphtheria toxin injection. (A) Serum aspartate transaminase (AST) activity levels over time after diphtheria toxin (DT) injection during the first 96 hours. DT doses of 0.5, 1, 1.5, 2, and 5 μg/kg were injected into 8 week-old Alb-TRECK/SCID mice, and serum AST activity was determined every 24 hours. Results are means ± standard error of the mean (n = 5/group). (B) Macroscopic views (left panels) and histology (hematoxylin and eosin (H&E) staining, right three panels) of mouse livers without and with administration of DT after 48 hours. DT dose: 1.5 μg/kg, serum AST activity of normal liver: 30 IU/L; serum AST activity of DT-treated liver: 12,000 IU/L. CV, central vein; PV, portal vein. Scale bars = 100 μm. (C) Kaplan–Meier survival curves of Alb-TRECK/SCID mice with different AST activities within 3 days after DT injection (numbers of mice in each group are indicated in the figure). *P < 0.05, **P < 0.01, ***P < 0.001. NS, not significant.
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Fig1: Alb-TRECK/SCID mice develop fulminant hepatic failure after diphtheria toxin injection. (A) Serum aspartate transaminase (AST) activity levels over time after diphtheria toxin (DT) injection during the first 96 hours. DT doses of 0.5, 1, 1.5, 2, and 5 μg/kg were injected into 8 week-old Alb-TRECK/SCID mice, and serum AST activity was determined every 24 hours. Results are means ± standard error of the mean (n = 5/group). (B) Macroscopic views (left panels) and histology (hematoxylin and eosin (H&E) staining, right three panels) of mouse livers without and with administration of DT after 48 hours. DT dose: 1.5 μg/kg, serum AST activity of normal liver: 30 IU/L; serum AST activity of DT-treated liver: 12,000 IU/L. CV, central vein; PV, portal vein. Scale bars = 100 μm. (C) Kaplan–Meier survival curves of Alb-TRECK/SCID mice with different AST activities within 3 days after DT injection (numbers of mice in each group are indicated in the figure). *P < 0.05, **P < 0.01, ***P < 0.001. NS, not significant.

Mentions: Serum AST activity reached a peak at 48 hours after DT injection and then returned toward basal levels by 96 hours (Figure 1A). This indicated that acute liver failure might have occurred and that the most severe liver damage might have been induced 48 hours after the administration of DT. All mice were dead within 48 hours after receiving 2 and 5 μg/kg of DT, and three mice were dead by 72 hours while two mice survived after administration of 1.5 μg/kg DT. All mice survived after administration of 0.5 and 1 μg/kg DT. Thus, we defined 1.5 μg/kg DT for 48 hours as a “sub-lethal dose” that could induce fulminant hepatic failure.Figure 1


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)

Alb-TRECK/SCID mice develop fulminant hepatic failure after diphtheria toxin injection. (A) Serum aspartate transaminase (AST) activity levels over time after diphtheria toxin (DT) injection during the first 96 hours. DT doses of 0.5, 1, 1.5, 2, and 5 μg/kg were injected into 8 week-old Alb-TRECK/SCID mice, and serum AST activity was determined every 24 hours. Results are means ± standard error of the mean (n = 5/group). (B) Macroscopic views (left panels) and histology (hematoxylin and eosin (H&E) staining, right three panels) of mouse livers without and with administration of DT after 48 hours. DT dose: 1.5 μg/kg, serum AST activity of normal liver: 30 IU/L; serum AST activity of DT-treated liver: 12,000 IU/L. CV, central vein; PV, portal vein. Scale bars = 100 μm. (C) Kaplan–Meier survival curves of Alb-TRECK/SCID mice with different AST activities within 3 days after DT injection (numbers of mice in each group are indicated in the figure). *P < 0.05, **P < 0.01, ***P < 0.001. NS, not significant.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig1: Alb-TRECK/SCID mice develop fulminant hepatic failure after diphtheria toxin injection. (A) Serum aspartate transaminase (AST) activity levels over time after diphtheria toxin (DT) injection during the first 96 hours. DT doses of 0.5, 1, 1.5, 2, and 5 μg/kg were injected into 8 week-old Alb-TRECK/SCID mice, and serum AST activity was determined every 24 hours. Results are means ± standard error of the mean (n = 5/group). (B) Macroscopic views (left panels) and histology (hematoxylin and eosin (H&E) staining, right three panels) of mouse livers without and with administration of DT after 48 hours. DT dose: 1.5 μg/kg, serum AST activity of normal liver: 30 IU/L; serum AST activity of DT-treated liver: 12,000 IU/L. CV, central vein; PV, portal vein. Scale bars = 100 μm. (C) Kaplan–Meier survival curves of Alb-TRECK/SCID mice with different AST activities within 3 days after DT injection (numbers of mice in each group are indicated in the figure). *P < 0.05, **P < 0.01, ***P < 0.001. NS, not significant.
Mentions: Serum AST activity reached a peak at 48 hours after DT injection and then returned toward basal levels by 96 hours (Figure 1A). This indicated that acute liver failure might have occurred and that the most severe liver damage might have been induced 48 hours after the administration of DT. All mice were dead within 48 hours after receiving 2 and 5 μg/kg of DT, and three mice were dead by 72 hours while two mice survived after administration of 1.5 μg/kg DT. All mice survived after administration of 0.5 and 1 μg/kg DT. Thus, we defined 1.5 μg/kg DT for 48 hours as a “sub-lethal dose” that could induce fulminant hepatic failure.Figure 1

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