Liver, Pancreas, and Biliary TractCorrection of liver disease by hepatocyte transplantation in a mouse model of progressive familial intrahepatic cholestasis☆,☆☆
Section snippets
Experimental protocol
Hepatocyte transplantation was performed in male mdr2(−/−) mice, the generation of which has been described previously.11 For donor mice, a transgenic A63 mouse strain that expresses the homologous human MDR3 gene behind an albumin promoter and was bred against the identical FVB background was used.13, 14 Compared with mdr2(+/+) control mice, phospholipid secretion in A63 mice is 180%.13, 14 MDR3-expressing hepatocytes were isolated from 3-month-old male A63 mice by ex vivo collagenase
Results
We tested the hypothesis that chronic damage to hepatocytes in a condition of defective biliary phospholipid secretion, i.e., in patients with PFIC type 3 and in mdr2(−/−) mice, causes a state of regenerative proliferation. This was done by immunostaining for Ki67 in samples from a PFIC type 3 patient and by staining for PCNA in sections from mdr2(−/−) livers (both markers stain nuclei of proliferating hepatocytes). Indeed, in livers from both the PFIC type 3 patients (Figure 1A) and mdr2(−/−)
Discussion
Lack of biliary phospholipid secretion leads to liver disease in both mice19 and humans.12 This is caused by the cytotoxic action of high concentrations of bile salts in the biliary tree. We hypothesized that this chronic damage to parenchymal cells leads to a continuous state of regenerative proliferation that makes these livers amenable to repopulation by transplanted hepatocytes. Our results now show that, at least for the mouse model, this hypothesis is correct. From the extensive
Acknowledgements
The authors thank Piet Borst and Bert Groen for critically reading the manuscript, Wilfried Meun for assistance with microphotography, Jos Mulder for assistance with PCNA immunohistochemistry, and Isabel Wesdorp for performing reverse-transcription polymerase chain reactions.
References (25)
- et al.
Intrahepatic transplantation of normal hepatocytes prevents Wilson's disease in Long-Evans cinnamon rats
Gastroenterology
(1996) - et al.
Homozygous disruption of the murine mdr2 P-glycoprotein gene leads to a complete absence of phospholipid from bile and to liver disease
Cell
(1993) - et al.
Effects of ursodeoxycholate and cholate feeding on liver disease in FVB mice with a disrupted mdr2 P-glycoprotein gene
Gastroenterology
(1996) - et al.
Liver regeneration
Science
(1997) Hepatic stem cells in liver regeneration
FASEB J
(1996)- et al.
Replacement of diseased mouse liver by hepatic cell transplantation
Science
(1994) - et al.
Hepatocytes corrected by gene therapy are selected in vivo in a murine model of hereditary tyrosinaemia type I
Nat Genet
(1996) - et al.
Selective repopulation of normal mouse liver by Fas/CD95-resistant hepatocytes
Nat Med
(1998) - et al.
Treatment of the Crigler-Najjar syndrome type I with hepatocyte transplantation
N Engl J Med
(1998) - et al.
Zonal regulation of gene expression during liver regeneration of urokinase transgenic mice
Hepatology
(1999)
Adenovirus-mediated gene therapy in a mouse model of hereditary tyrosinemia type I
Hum Gene Ther
Therapeutic trials in the murine model of hereditary tyrosinaemia type I: a progress report
J Inherit Metab Dis
Cited by (0)
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Address requests for reprints to: Ronald P. J. Oude Elferink, Ph.D., Department of Gastroenterology and Hepatology Fo-116, Meibergdreef 9, 1105 AZ Amsterdam, The Netherlands. e-mail: [email protected]; fax: (31) 20-5669190.
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Supported by project no. 902-23-097 from the Dutch Association for Scientific Research.