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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

Functional assessments of humanized livers in Alb-TRECK/SCID mice. (A) Repopulation rates of human primary fetal liver cell (FLC)- and human hepatic stem cell (HpSC)-derived human hepatocytes in Alb-TRECK/SCID mice at about 8 weeks after transplantation. Results are means ± standard error of the mean (n = 12 and 14, respectively). (B) Serum human albumin (ALB) levels in mice with humanized livers derived from human primary FLCs and HpSCs at about 7 to 8 weeks after transplantation. Results are means ± standard error of the mean (n = 6/group). (C) Human-specific ketoprofen (KTP; CYP2C9) drug biotransformation in humanized Alb-TRECK/SCID mice at about 8 weeks after transplantation. (D) Analysis of human-specific CYP2D6-mediated debrisoquine (DEB) metabolism in human HpSC-derived humanized Alb-TRECK/SCID mice by metabolic ratios of the metabolite 4-hyroxydebrisoqune (4OH-DEB) to DEB. Results are means ± standard error of the mean (n ≥ 4/group). **P < 0.01, ***P < 0.001. ND, not detectable; NS, not significant; Sham, mice transplanted with saline.
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Fig6: Functional assessments of humanized livers in Alb-TRECK/SCID mice. (A) Repopulation rates of human primary fetal liver cell (FLC)- and human hepatic stem cell (HpSC)-derived human hepatocytes in Alb-TRECK/SCID mice at about 8 weeks after transplantation. Results are means ± standard error of the mean (n = 12 and 14, respectively). (B) Serum human albumin (ALB) levels in mice with humanized livers derived from human primary FLCs and HpSCs at about 7 to 8 weeks after transplantation. Results are means ± standard error of the mean (n = 6/group). (C) Human-specific ketoprofen (KTP; CYP2C9) drug biotransformation in humanized Alb-TRECK/SCID mice at about 8 weeks after transplantation. (D) Analysis of human-specific CYP2D6-mediated debrisoquine (DEB) metabolism in human HpSC-derived humanized Alb-TRECK/SCID mice by metabolic ratios of the metabolite 4-hyroxydebrisoqune (4OH-DEB) to DEB. Results are means ± standard error of the mean (n ≥ 4/group). **P < 0.01, ***P < 0.001. ND, not detectable; NS, not significant; Sham, mice transplanted with saline.

Mentions: At about 8 weeks after transplantation, the level of liver repopulation with human donor cells and human ALB concentrations in mouse sera were determined. The average liver repopulation rate for human primary FLC- and HpSC-derived humanized livers were 76% and 71%, respectively; no significant difference was observed. Several humanized livers reached liver repopulation levels of about 100%, which indicated that almost the entire mouse liver had been reconstituted with human hepatocytes (Figure 6A). Humanized livers derived from human HpSCs resulted in more human ALB secretion than those from human primary FLCs, and no human ALB could be detected in mice after saline transplantation (Figure 6B). The drug metabolism profiles based on gene expression and human ALB secretion patterns suggested the potential of humanized livers derived from human HpSCs for early identification of major drug metabolites in vivo.Figure 6


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)

Functional assessments of humanized livers in Alb-TRECK/SCID mice. (A) Repopulation rates of human primary fetal liver cell (FLC)- and human hepatic stem cell (HpSC)-derived human hepatocytes in Alb-TRECK/SCID mice at about 8 weeks after transplantation. Results are means ± standard error of the mean (n = 12 and 14, respectively). (B) Serum human albumin (ALB) levels in mice with humanized livers derived from human primary FLCs and HpSCs at about 7 to 8 weeks after transplantation. Results are means ± standard error of the mean (n = 6/group). (C) Human-specific ketoprofen (KTP; CYP2C9) drug biotransformation in humanized Alb-TRECK/SCID mice at about 8 weeks after transplantation. (D) Analysis of human-specific CYP2D6-mediated debrisoquine (DEB) metabolism in human HpSC-derived humanized Alb-TRECK/SCID mice by metabolic ratios of the metabolite 4-hyroxydebrisoqune (4OH-DEB) to DEB. Results are means ± standard error of the mean (n ≥ 4/group). **P < 0.01, ***P < 0.001. ND, not detectable; NS, not significant; Sham, mice transplanted with saline.
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Fig6: Functional assessments of humanized livers in Alb-TRECK/SCID mice. (A) Repopulation rates of human primary fetal liver cell (FLC)- and human hepatic stem cell (HpSC)-derived human hepatocytes in Alb-TRECK/SCID mice at about 8 weeks after transplantation. Results are means ± standard error of the mean (n = 12 and 14, respectively). (B) Serum human albumin (ALB) levels in mice with humanized livers derived from human primary FLCs and HpSCs at about 7 to 8 weeks after transplantation. Results are means ± standard error of the mean (n = 6/group). (C) Human-specific ketoprofen (KTP; CYP2C9) drug biotransformation in humanized Alb-TRECK/SCID mice at about 8 weeks after transplantation. (D) Analysis of human-specific CYP2D6-mediated debrisoquine (DEB) metabolism in human HpSC-derived humanized Alb-TRECK/SCID mice by metabolic ratios of the metabolite 4-hyroxydebrisoqune (4OH-DEB) to DEB. Results are means ± standard error of the mean (n ≥ 4/group). **P < 0.01, ***P < 0.001. ND, not detectable; NS, not significant; Sham, mice transplanted with saline.
Mentions: At about 8 weeks after transplantation, the level of liver repopulation with human donor cells and human ALB concentrations in mouse sera were determined. The average liver repopulation rate for human primary FLC- and HpSC-derived humanized livers were 76% and 71%, respectively; no significant difference was observed. Several humanized livers reached liver repopulation levels of about 100%, which indicated that almost the entire mouse liver had been reconstituted with human hepatocytes (Figure 6A). Humanized livers derived from human HpSCs resulted in more human ALB secretion than those from human primary FLCs, and no human ALB could be detected in mice after saline transplantation (Figure 6B). The drug metabolism profiles based on gene expression and human ALB secretion patterns suggested the potential of humanized livers derived from human HpSCs for early identification of major drug metabolites in vivo.Figure 6

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