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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

Characterization of human primary fetal liver cell- and human hepatic stem cell-derived human hepatocytes in Alb-TRECK/SCID mice. (A) Immunohistochemistry to distinguish between human hepatocytes stained with anti-human CK8/18 (green) antigen and anti-human nuclear antigen (aqua blue) in human primary fetal liver cell- (FLC; upper panels) and human hepatic stem cell- (HpSC; lower panels) derived humanized livers at 6 weeks after transplantation. (B) Immunohistochemistry analyses for human albumin, human CK19, and human CK8/18 expression in human primary FLC- (upper panels) and HpSC- (lower panels) derived livers at 6 weeks after transplantation. White dashed line: mouse liver region distinguished from human liver region. Nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI, blue). Scale bars = 100 μm. m, mouse liver region; h, human donor cell-derived human region.
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Fig4: Characterization of human primary fetal liver cell- and human hepatic stem cell-derived human hepatocytes in Alb-TRECK/SCID mice. (A) Immunohistochemistry to distinguish between human hepatocytes stained with anti-human CK8/18 (green) antigen and anti-human nuclear antigen (aqua blue) in human primary fetal liver cell- (FLC; upper panels) and human hepatic stem cell- (HpSC; lower panels) derived humanized livers at 6 weeks after transplantation. (B) Immunohistochemistry analyses for human albumin, human CK19, and human CK8/18 expression in human primary FLC- (upper panels) and HpSC- (lower panels) derived livers at 6 weeks after transplantation. White dashed line: mouse liver region distinguished from human liver region. Nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI, blue). Scale bars = 100 μm. m, mouse liver region; h, human donor cell-derived human region.

Mentions: Prior to testing whether the transplanted human immature hepatocytes in mice had the potential for differentiation and be functional in vivo, we firstly confirmed human hepatocytes existed in mouse livers by immunohistochemical analysis at about 6 weeks after transplanting human primary FLCs (Figure 4A, left panels) or HpSCs (Figure 4B, left panels). Liver sections from these mice that were specifically positively co-stained with human nuclei and human cytokeratin 8/18 (CK8/18) antibodies were donor cell-derived human hepatocytes, while the original liver regions in mice were negative for these markers.Figure 4


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)

Characterization of human primary fetal liver cell- and human hepatic stem cell-derived human hepatocytes in Alb-TRECK/SCID mice. (A) Immunohistochemistry to distinguish between human hepatocytes stained with anti-human CK8/18 (green) antigen and anti-human nuclear antigen (aqua blue) in human primary fetal liver cell- (FLC; upper panels) and human hepatic stem cell- (HpSC; lower panels) derived humanized livers at 6 weeks after transplantation. (B) Immunohistochemistry analyses for human albumin, human CK19, and human CK8/18 expression in human primary FLC- (upper panels) and HpSC- (lower panels) derived livers at 6 weeks after transplantation. White dashed line: mouse liver region distinguished from human liver region. Nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI, blue). Scale bars = 100 μm. m, mouse liver region; h, human donor cell-derived human region.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig4: Characterization of human primary fetal liver cell- and human hepatic stem cell-derived human hepatocytes in Alb-TRECK/SCID mice. (A) Immunohistochemistry to distinguish between human hepatocytes stained with anti-human CK8/18 (green) antigen and anti-human nuclear antigen (aqua blue) in human primary fetal liver cell- (FLC; upper panels) and human hepatic stem cell- (HpSC; lower panels) derived humanized livers at 6 weeks after transplantation. (B) Immunohistochemistry analyses for human albumin, human CK19, and human CK8/18 expression in human primary FLC- (upper panels) and HpSC- (lower panels) derived livers at 6 weeks after transplantation. White dashed line: mouse liver region distinguished from human liver region. Nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI, blue). Scale bars = 100 μm. m, mouse liver region; h, human donor cell-derived human region.
Mentions: Prior to testing whether the transplanted human immature hepatocytes in mice had the potential for differentiation and be functional in vivo, we firstly confirmed human hepatocytes existed in mouse livers by immunohistochemical analysis at about 6 weeks after transplanting human primary FLCs (Figure 4A, left panels) or HpSCs (Figure 4B, left panels). Liver sections from these mice that were specifically positively co-stained with human nuclei and human cytokeratin 8/18 (CK8/18) antibodies were donor cell-derived human hepatocytes, while the original liver regions in mice were negative for these markers.Figure 4

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