Hepatoblasts and method of isolating same

Abstract

This invention relates to methods of isolating hepatoblasts utilizing panning techniques and fluorescence activated cell sorting. This invention further relates to isolated hepatoblasts and to a method of treating liver dysfunction as well as to methods of forming artificial livers.

1 - 34 . (canceled)  
   
   
       35 . A method of providing mature hepatocytes to a subject in need thereof comprising parenterally administering to the subject a population of isolated hepatoblasts, the population having been obtained by: 
 (a) preparing a single cell suspension of liver cells;    (b) panning said suspension utilizing antibodies specific for mature hepatocytes, mature bile duct cells, endothelial cells, mesenchymal cells, and hemopoietic cells so as to remove mature hepatocytes, mature bile duct cells, endothelial cells, mesenchymal cells, and hemopoietic cells from said suspension;    (c) performing fluorescence activated cell sorting utilizing said antibodies so as to remove remaining mature hepatocytes, mature bile duct cells, endothelial cells, mesenchymal cells, and hemopoietic cells from said suspension; and    (d) performing multiparametric fluorescence activated cell sorting on said suspension utilizing at least one antibody to a hepatic cell marker, side scatter, forward scatter and/or autofluorescence to isolate said hepatic progenitors; and    in which the hepatoblasts differentiate into mature hepatocytes in vivo.    
   
   
       36 . The method of  claim 35  in which the hepatoblasts are isolated from embryonic or neonatal liver.  
   
   
       37 . The method of  claim 35  in which the hepatoblasts are isolated from adult human liver.  
   
   
       38 . The method of  claim 35  in which the subject in need has a liver dysfunction.  
   
   
       39 . The method of  claim 35  in which the administering comprises injecting, transplanting or grafting.  
   
   
       40 . The method of  claim 39  in which the injecting is into the spleen, portal venous system, bile duct or peritoneum of the subject.  
   
   
       41 . The method of  claim 35  in which the population of isolated hepatoblasts is obtained from a donor.