Volume 122, Issue 2, Pages 438-447 (February 2002) Early cell transplantation in LEC rats modeling Wilson's disease eliminates hepatic copper with reversal of liver disease  Harmeet Malhi, Adil N. Irani, Irene Volenberg, Michael L. Schilsky, Sanjeev Gupta  Gastroenterology  Volume 122, Issue 2, Pages 438-447 (February 2002) DOI: 10.1053/gast.2002.31086 Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 1 Analysis of animals 5 months after cell transplantation. (A) Bile copper excretion is shown in 3 LEC rats in the basal state and after stimulation with a copper-histidine bolus. The range of bile copper excretion in these conditions is shown in LEA and LEC rats. After a copper bolus, LEA rats show up to 2- ± 1-fold greater copper excretion within 30 minutes. Although basal copper excretion increased in 1 treated LEC rat (P5-3), copper stimulation did not elicit a further increase in copper excretion. (B) Liver from rat P5-1 (a) showing a periportal area with extensive abnormalities, including polyploid cells containing megalonuclei (arrows), oval cell activation and cholangiofibrosis (arrowhead), steatosis, and mitotic activity (inset, arrow). (b) Rat P5-3, with a portal area showing healthy hepatocytes containing nuclei of normal size without fatty or hydropic change. (C) Additional data 5 months after cell transplantation. Gastroenterology 2002 122, 438-447DOI: (10.1053/gast.2002.31086) Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 2 atp7b mRNA expression. (A) Gel electrophoresis analysis of polymerase chain reaction products with a 380-base pair atp7b band and a 188-base pair β-actin band. Lanes 1 and 20 contain molecular-weight markers, lane 2 was a negative control with polymerase chain reaction mix alone, lanes 3 and 4 contained samples from LEA rat livers, lane 5 contains a sample from LEC rat liver, and lanes 6–19 contain samples from various cell recipients. Arrowheads indicate the presence of atp7b band. (B) The identity of various animals and densitometric scanning data indicating the proportion of liver repopulated. Gastroenterology 2002 122, 438-447DOI: (10.1053/gast.2002.31086) Copyright © 2002 American Gastroenterological Association Terms and Conditions

Fig. 3 Representative histologic findings in liver samples obtained 20 months after cell transplantation. (A) Liver from an untreated LEC rat showing extensive cholangiofibrosis in a portal area (P). Inset, polyploid change with megalonuclei (arrow). (B) Liver from a LEA rat with no specific histologic abnormalities. (C) Liver from a partially responsive cell recipient (C3.1) with atp7b mRNA expression and a significant decrease but not normalization in liver copper level. Histology was markedly improved without biliary abnormalities, although scattered hepatocytes showed polyploid change (inset, arrow), steatosis, or hydropic changes, especially in perivenous areas. (D) Liver from a fully responsive LEC rat (C4.0) showing normal histology with no difference from the histology observed in normal LEA rats. (H&E stain; original magnifications: A, 100×; B-D, 200×; insets, 400×.). Gastroenterology 2002 122, 438-447DOI: (10.1053/gast.2002.31086) Copyright © 2002 American Gastroenterological Association Terms and Conditions