Oligopeptide-free cell culture media

Abstract

The present invention relates to oligopeptide-free cell culture media comprising at least 0.5 mg/L of a polyamine and to methods for cultivating cells in said oligopeptide-free cell culture media comprising at least 0.5 mg/L of a polyamine. The invention also relates to methods for expressing at least one protein in a medium comprising at least 0.5 mg/L of a polyamine and to methods for producing at least one virus in a medium comprising at least 0.5 mg/L of a polyamine.

1 . An oligopeptide-free cell culture medium, comprising at least 0.5 mg/L of a polyamine.  
     
     
         2 . The oligopeptide-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 combinations thereof.  
     
     
         3 . The oligopeptide-free cell culture medium according to  claim 1 , wherein the polyamine is synthetically produced.  
     
     
         4 . The oligopeptide-free cell culture medium according to  claim 1 , wherein the polyamine is present in the culture in a concentration ranging from about 0.5 to about 30 mg/L.  
     
     
         5 . The oligopeptide-free cell culture medium according to  claim 1 , wherein the does not comprise oligopeptides having twenty or more amino acids.  
     
     
         6 . The oligopeptide-free cell culture medium according to  claim 1 , wherein the medium does not comprise oligopeptides having three or more amino acids, optionally comprising glutathione.  
     
     
         7 . The oligopeptide-free cell culture medium according to  claim 1 , wherein the medium does not comprise oligopeptides having two or more amino acids, optionally comprising glutathione and/or at least one stable form of glutamine.  
     
     
         8 . The oligopeptide-free cell culture medium according to  claim 1 , wherein the medium is chemically defined.  
     
     
         9 . A method for cultivating cells, comprising the steps of: 
 (a) providing an oligopeptide-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 . A method for expressing at least one protein, comprising the steps of: 
 (a) providing a culture of cells;    (b) introducing at least one nucleic acid sequence comprising a sequence coding for at least one protein into the cells;    (c) selecting the cells carrying the nucleic acid sequence; and    (d) expressing the protein in the cells in a comprising at least 0.5 mg/L of a polyamine.    
     
     
         12 . The method according to  claim 11 , wherein the medium is an oligo-peptide free medium according to  claim 1 .  
     
     
         13 . The method according to  claim 11 , wherein the protein is selected from the group of coagulation factor VII, coagulation factor VIII, coagulation factor IX, vWF, ADAMTS13, and furin.  
     
     
         14 . The method according to  claim 11 , wherein the cells are CHO cells, 293 cells or BHK cells.  
     
     
         15 . The method according to  claim 11 , wherein the cell/protein combination is selected from the group consisting of CHO cells/coagulation factor VIII, CHO cells/coagulation factor VII, CHO cells/ADAMTS13, CHO cells/furin, 293 cells/coagulation factor IX.  
     
     
         16 . A method for producing at least one virus, comprising the steps of: 
 (a) providing a culture of cells;    (b) infecting the cells with the at least one virus;    (c) selecting the virus-infected cells; and    (d) propagating the at least one virus in the cells in a medium comprising at least 0.5 mg/L of a polyamine.    
     
     
         17 . The method according to  claim 16 , wherein the is an oligo-peptide free medium according to  claim 1 .  
     
     
         18 . The method according to  claim 16 , wherein the virus is selected from the group of poxviruses, coronaviruses, orthomyxoviruses, paramyxoviruses, retroviruses, togaviruses, flaviviruses, enteroviruses, picornaviruses, arenaviruses, herpesviruses, and adenoviruses.  
     
     
         19 . The method according to  claim 16 , wherein the virus is selected from the group of vaccinia virus, SARS virus, influenza A virus, influenza B virus, Lenti virus, Ross River virus, West Nile virus, Yellow Fever virus, FSME virus, and hepatitis A virus.  
     
     
         20 . The method according to  claim 16 , wherein the cells are Vero cells or chicken embryo cells.  
     
     
         21 . The method according to  claim 16 , wherein the cells/virus combination is selected from the group consisting of Vero cells/attenuated vaccinia, Vero cells/vaccinia, Vero cells/hepatitis A, Vero cells/influenza virus, Vero cells/West Nile virus, Vero cells/SARS virus, Vero cells/Yellow Fever virus, chicken embryo cells/FSME virus, and chicken embryo cells/Modified Vaccinia Virus Ankara (MVA).