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AssigneeLife Technologies Corporation
InventorsStephen F. Gorfien, Richard M. Fike, Glenn P. Godwin, Joyce L. Dzimian, David A. Epstein, Dale Gruber, Don McClure, Paul J. Price
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The present invention provides a cell culture medium formulation that supports the in vitro cultivation, particularly in suspension, of mammalian cells, particularly epithelial cells and fibroblast cells, and methods for cultivating mammalian cells in suspension in vitro using these media. The media comprise a basal medium and a polyanionic or polyanionic compound, preferably a polysulfonated or polysulfated compound, and more preferably dextran sulfate. The present invention also provides chemically defined, protein-free eukaryotic cell culture media comprising an iron chelate and zinc, which is capable of supporting the growth (and particularly the high-density growth of mammalian cells) in suspension culture, increasing the level of expression of recombinant protein in cultured cells, and/or increasing virus production in cultured cells.
A method for producing a recombinant protein in vitro, comprising culturing a recombinant CHO cell in suspension, in a serum-free and protein-free culture medium comprising zinc at a final concentration of about 0.00007 to 0.00073 g/L, and iron at a final concentration of about 0.00028 to 0.011 g/L, under conditions suitable for supporting the cultivation of the CHO cell to a density of about 5×106 cells/ml, in said suspension and expression of said recombinant protein, the conditions comprising culturing the CHO cell at about 37 degrees Celsius in a humidified atmosphere containing about 3-20% carbon dioxide in air, and wherein said serum-free and protein-free culture medium supports increased levels of cell growth as compared to culture in FMX-8 medium.
Dependencies are inferred from the claim text.