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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

Expressions of human hepatocyte-related genes in human primary fetal liver cell- and human hepatic stem cell-derived humanized livers. (A) Results of quantitative PCR for the expression of functional hepatocyte markers, including hALB, hAFP, hCYP3A4, hCYP2C9, and hCYP2C19, in samples taken before and about 8 weeks after human primary fetal liver cell (FLC) and human hepatic stem cell (HpSC) transplantation. Results are mean ± standard error of the mean (n ≥ 4 mice/group). (B) Heat maps for 83 human-specific drug metabolism genes and transcription factors, shown separately for independent experiments to analyze human primary FLCs and HpSCs before and at 8 weeks after cell transplantation. Human adult hepatocytes were used as a positive control. AH, adult hepatocyte; ND, not detectable; Sham, mice transplanted with saline.
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Fig5: Expressions of human hepatocyte-related genes in human primary fetal liver cell- and human hepatic stem cell-derived humanized livers. (A) Results of quantitative PCR for the expression of functional hepatocyte markers, including hALB, hAFP, hCYP3A4, hCYP2C9, and hCYP2C19, in samples taken before and about 8 weeks after human primary fetal liver cell (FLC) and human hepatic stem cell (HpSC) transplantation. Results are mean ± standard error of the mean (n ≥ 4 mice/group). (B) Heat maps for 83 human-specific drug metabolism genes and transcription factors, shown separately for independent experiments to analyze human primary FLCs and HpSCs before and at 8 weeks after cell transplantation. Human adult hepatocytes were used as a positive control. AH, adult hepatocyte; ND, not detectable; Sham, mice transplanted with saline.

Mentions: We also evaluated human drug metabolism-related gene expression by quantitative PCR and microarray analysis to assess whether human primary FLC- and HpSC-derived humanized livers could be used for human drug metabolism studies. At 8 weeks after transplantation, we observed very high gene expression levels associated with the human hepatic functional markers ALB, AFP, and cytochrome P450, including CYP3A4, 2C9, and 2C19, which collectively metabolize over 80% of clinical drugs (Figure 5A). Almost none of the probes on the human gene expression array had cross-hybridized with murine mRNA.Figure 5


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)

Expressions of human hepatocyte-related genes in human primary fetal liver cell- and human hepatic stem cell-derived humanized livers. (A) Results of quantitative PCR for the expression of functional hepatocyte markers, including hALB, hAFP, hCYP3A4, hCYP2C9, and hCYP2C19, in samples taken before and about 8 weeks after human primary fetal liver cell (FLC) and human hepatic stem cell (HpSC) transplantation. Results are mean ± standard error of the mean (n ≥ 4 mice/group). (B) Heat maps for 83 human-specific drug metabolism genes and transcription factors, shown separately for independent experiments to analyze human primary FLCs and HpSCs before and at 8 weeks after cell transplantation. Human adult hepatocytes were used as a positive control. AH, adult hepatocyte; ND, not detectable; Sham, mice transplanted with saline.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig5: Expressions of human hepatocyte-related genes in human primary fetal liver cell- and human hepatic stem cell-derived humanized livers. (A) Results of quantitative PCR for the expression of functional hepatocyte markers, including hALB, hAFP, hCYP3A4, hCYP2C9, and hCYP2C19, in samples taken before and about 8 weeks after human primary fetal liver cell (FLC) and human hepatic stem cell (HpSC) transplantation. Results are mean ± standard error of the mean (n ≥ 4 mice/group). (B) Heat maps for 83 human-specific drug metabolism genes and transcription factors, shown separately for independent experiments to analyze human primary FLCs and HpSCs before and at 8 weeks after cell transplantation. Human adult hepatocytes were used as a positive control. AH, adult hepatocyte; ND, not detectable; Sham, mice transplanted with saline.
Mentions: We also evaluated human drug metabolism-related gene expression by quantitative PCR and microarray analysis to assess whether human primary FLC- and HpSC-derived humanized livers could be used for human drug metabolism studies. At 8 weeks after transplantation, we observed very high gene expression levels associated with the human hepatic functional markers ALB, AFP, and cytochrome P450, including CYP3A4, 2C9, and 2C19, which collectively metabolize over 80% of clinical drugs (Figure 5A). Almost none of the probes on the human gene expression array had cross-hybridized with murine mRNA.Figure 5

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