Animal protein-free media for cultivation of cells

Abstract

The present invention relates to animal protein-free cell culture media comprising polyamines and a plant and/or yeast-derived hydrolysate. The invention also relates to animal protein-free culturing processes, wherein cells can be cultivated, propagated and passaged without adding supplementary animal proteins in the culture medium. These processes are useful in cultivating cells, such as recombinant cells or cells infected with a virus, and for producing biological products by cell culture processes.

1 . An animal protein-free cell culture medium, comprising at least one polyamine and at least one protein hydrolysate derived from the group consisting of plants and yeast.  
     
     
         2 . The animal protein-free cell culture medium according to  claim 1 , wherein the polyamine is present in the culture medium in a concentration ranging from about 0.5 to about 10 mg/L.  
     
     
         3 . The animal protein-free cell culture medium according to  claim 1 , wherein the polyamine is selected from the group consisting of cadaverine, putrescine, spermidine, spermine, agmatine, ornithine, and a combination thereof.  
     
     
         4 . The animal protein-free cell culture medium according to  claim 1 , wherein the polyamine is putrescine in a concentration ranging from about 0.5 to about 10 mg/L, and the protein hydrolysate is soy hydrolysate in a concentration ranging from about 0.05% (w/v) to about 5% (w/v)  
     
     
         5 . The animal protein-free cell culture medium according to  claim 1 , wherein the polyamine originates from a source other than a protein hydrolysate.  
     
     
         6 . The animal protein-free cell culture medium according to  claim 1 , wherein the polyamine is present in the culture medium in a concentration ranging from about 0.5 to 30 mg/L.  
     
     
         7 . The animal protein-free cell culture medium according to  claim 1 , wherein the protein hydrolysate is present in the culture medium in a total concentration ranging from about 0.05% (w/v) to about 5% (w/v) for all protein hydrolysates.  
     
     
         8 . The animal protein-free cell culture medium according to  claim 1 , wherein the protein hydrolysate is derived from a plant selected from the group consisting cereals and soy.  
     
     
         9 . A method for cultivating cells, comprising the steps of: 
 (a) providing an animal protein-free cell culture medium according to  claim 1 , and    (b) propagating the cells in the medium to form a cell culture.    
     
     
         10 . The method according to  claim 9 , wherein the cells are selected from the group consisting of mammalian cells, insect cells, avian cells, bacterial cells, and yeast cells.  
     
     
         11 . The method according to  claim 9 , wherein the cells are cultivated by a method selected from the group consisting of batch-cultivation, feed-batch-cultivation, perfusion cultivation, and chemostat-cultivation.  
     
     
         12 . A method for expressing a target protein, comprising the steps of: 
 a) providing a culture of cells that have been grown in an animal protein-free cell culture medium according to  claim 1;     b) introducing a nucleic acid sequence comprising a sequence coding for the target protein into the cells;    c) selecting the cells carrying the nucleic acid sequence; and    d) selectively inducing the expression of the target protein in the cells.    
     
     
         13 . The method according to  claim 12 , wherein the cells are selected from the group consisting of mammalian cells, insect cells, avian cells, bacterial cells, and yeast cells.  
     
     
         14 . The method according to  claim 1  wherein the cell/target protein combination is selected from the group consisting of CHO cells/coagulation factor VIII, BHK cells/erythropoietin, Epstein Barr virus transformed, immortalized human B cells/human antibodies.  
     
     
         15 . The method according to  claim 12 , wherein the cells are cultivated by a method selected from the group consisting of batch-cultivation, feed-batch-cultivation, perfusion cultivation, and chemostat-cultivation.  
     
     
         16 . A method for producing a virus, comprising the steps of: 
 a) providing a culture of cells that have been grown in an animal protein-free cell culture medium according to  claim 1;     b) infecting the cells with-the virus;    c) selecting the virus-infected cells; and    d) incubating the cells to propagate the virus.    
     
     
         17 . The method according to  claim 16 , wherein the cells are selected from the group consisting of mammalian cells, insect cells, avian cells, bacterial cells, and yeast cells.  
     
     
         18 . The method according to  claim 16 , wherein the cell/virus combination is selected from the group consisting of Vero cell/attenuated vaccinia, Vero cell/vaccinia, Vero cell/hepatitis A, Vero cell/influenza virus, Vero cell/West Nile virus, Vero cell/SARS virus, and chicken embryo cells/FSME virus.  
     
     
         19 . The method according to  claim 16 , wherein the cells are cultivated by a method selected from the group consisting of batch-cultivation, feed-batch-cultivation, perfusion cultivation, and chemostat-cultivation.