Self-renewing pluripotent hepatic stem cells

Abstract

The present invention provides clonal pluripotent hepatic stem cells using flow cytometry and in vitro single-cell-based assays. These cells possess multilineage differentiation potential and self-renewing capability. These cells could be clonally propagated in culture, where they continuously produced hepatocytes and cholangiocytes as descendants while maintaining primitive stem cells. When cells that expanded in vitro were transplanted into recipient animals, they morphologically and functionally differentiated into hepatocytes and cholangiocytes, with reconstitution of hepatocyte and bile duct structures. Furthermore, these cells differentiated into pancreatic ductal and acinar cells or intestinal epithelial cells when transplanted into pancreas or duodenal wall. These data indicate that self-renewing multipotent stem cells persist in the developing mouse liver and that such cells can be induced to become cells of other organs of endodermal origin under appropriate microenvironment. Manipulation of hepatic stem cells may provide new insight into therapies for diseases of the digestive system.

1 . A cloned, self-renewing pluripotent hepatic stem cell or progeny thereof.  
     
     
         2 . The stem cell according to  claim 1  which is from primate.  
     
     
         3 . The stem cell according to  claim 1  which is human.  
     
     
         4 . The stem cell according to  claim 1 , which is capable of differentiating into hepatocytes or cholangiocytes.  
     
     
         5 . The stem cell according to  claim 1 , which is capable of differentiating into pancreatic ductal, acinar, or intestinal epithelial cells.  
     
     
         6 . A purified composition essentially consisting of cloned, self-renewing pluripotent hepatic stem cells according to  claim 1 .  
     
     
         7 . A method for producing cloned, self-renewing pluripotent stem cells, comprising: 
 providing cells; and    fractionating said cells by selecting cells expressing c-Met +  CD49f +/low .    
     
     
         8 . The method according to  claim 7 , said step of fractionating further comprising selecting cells expressing c-Kit −  CD45 −  TER119 − .  
     
     
         9 . The method according to  claim 7 , wherein said cell is a liver cell.  
     
     
         10 . The method according to  claim 7 , said cell is fetal.  
     
     
         11 . A differentiated cell derived from a cloned, self-renewing pluripotent hepatic stem cell, or progeny thereof.  
     
     
         12 . The differentiated cell according to  claim 11 , which is selected from the group consisting of a hepatocyte, a cholangiocyte, a pancreatic ductal cell, an acinar cell, and an intestinal epithelial cell.  
     
     
         13 . A tissue regenerated from a cloned, self-renewing pluripotent hepatic stem cell, or progeny thereof.  
     
     
         14 . The tissue according to  claim 13  which is a liver tissue.  
     
     
         15 . An organ regenerated from a cloned, self-renewing pluripotent hepatic stem cell, or progeny thereof.  
     
     
         16 . The organ according to  claim 15  which is a liver.  
     
     
         17 . A method of transplanting cells into a recipient host, comprising: 
 obtaining a cloned, self-renewing pluripotent hepatic stem cell, or progeny thereof; and    transplanting said stem cell into the host.    
     
     
         18 . A method of transplanting a tissue into a recipient host, comprising: 
 obtaining a cloned, self-renewing pluripotent hepatic stem cell, or progeny thereof;    regenerating a tissue from the stem cell; and    transplanting said tissue into the host.    
     
     
         19 . The method according to  claim 18 , wherein said tissue is a liver tissue.  
     
     
         20 . A method of transplanting an organ into a recipient host, comprising: 
 obtaining a cloned, self-renewing pluripotent hepatic stem cell, or progeny thereof;    regenerating an organ from the stem cell; and transplanting said organ into the host.    
     
     
         21 . The method according to  claim 20 , wherein said organ is a liver.