Cell culture media for mammalian cells

Abstract

A chemically defined mammalian cell culture medium is provided that supports maintenance and long term clonal growth of mammalian hepatocytes and other cells.

1 . A method of producing recombinant pancreatic islet cells expressing a heterologous gene comprising: 
 producing pancreatic islet cells in vitro without serum by introducing pancreatic islet cells into a cell culture medium in vitro, said culture medium comprising 1-150 mg/L arginine; 1-120 mg/L proline; 1-3050 mg/L nicotinamide; 0.1-100 mg/L transferrin chelated with iron; greater than 10 11  M insulin or insulin-like growth factors; 10 -12  M-10 -3  M glucocorticoid steroid; 1-6000 μg/L zinc salt; 1-250 μg/L manganese salt; 1-1000 μg/L copper salt; 1-150 μg/L selenium salt; 2.0-10.0 mM L-glutamine; 0.01-5.0 g/L D-galactose or 0.01-5.0 g/L D-glucose, or when both D-galactose and D-glucose are included together, 0.01-8.0 g/L, and culturing said introduced cells in said medium; and    transforming said cultured pancreatic islet cells with a nucleic acid capable of expressing said gene in said cells.    
   
   
       2 . A method of using pancreatic islet cells produced according to  claim 1  comprising infusing said pancreatic islet cells into a patient and allowing said gene to be expressed.  
   
   
       3 . A method for pancreatic islet cell transplantation comprising introducing recombinant pancreatic islet cells produced according to  claim 1  into a patient.  
   
   
       4 . A method for manufacturing a gene product comprising culturing recombinant pancreatic islet cells produced according to  claim 1  and recovering the gene product.  
   
   
       5 . A method for testing a drug comprising introducing said drug to recombinant pancreatic islet cells produced according to  claim 1  and assaying for the effect of the drug.  
   
   
       6 . A method of producing recombinant hepatocytes expressing a heterologous gene comprising: 
 producing hepatocytes in vitro without serum by introducing hepatocytes into a cell culture medium in vitro, said culture medium comprising 1-150 mg/L arginine; 1-120 mg/L proline; 1-3050 mg/L nicotinamide; 0.1-100 mg/L transferrin chelated with iron; greater than 10 -11  M insulin or insulin-like growth factors; 10 -12  M-10 -3  M glucocorticoid steroid; 1-6000 μg/L zinc salt; 1-250 μg/L manganese salt; 1-1000 μg/L copper salt; 1-150 μg/L selenium salt; 2.0-10.0 mM L-glutamine; 0.01-5.0 g/L D-galactose or 0.01-5.0 g/L D-glucose, or when both D-galactose and D-glucose are included together, 0.01-8.0 g/L, and culturing said introduced cells in said medium; and    transforming said cultured hepatocytes with a nucleic acid capable of expressing said gene in said cells.    
   
   
       7 . A method of using hepatocytes produced according to  claim 6  comprising infusing said hepatocytes into a patient and allowing said gene to be expressed.  
   
   
       8 . A method for hepatocytes transplantation comprising introducing recombinant hepatocytes produced according to  claim 6  into a patient.  
   
   
       9 . A method for manufacturing a gene product comprising culturing recombinant hepatocytes produced according to  claim 6  and recovering the gene product.  
   
   
       10 . A method for testing a drug comprising introducing said drug to recombinant hepatocytes produced according to  claim 6  and assaying for the effect of the drug.