Production of viruses in tissue culture without use of serum
AssigneeThe Dow Chemical Company
InventorsStefan A. Weiss, Harry L. Torney, William J. Caldwell
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Abstract
Growth of cells in tissue culture and virus vaccine production in tissue culture in the absence of serum are disclosed. The cells in tissue culture are grown initially in a chemically defined medium containing protamine zinc insulin, and the protamine zinc insulin medium is removed before virus propagation. Supplementation with serum or serum fractions can be eliminated entirely; and improved virus production in a serum free system can be obtained.
Claims15
In a method which comprises inoculating a chemically defined tissue culture medium with living animal cells and incubating the cells and medium in tissure culture until significant cell growth is obtained, and inoculating the cells with virus, the improvement wherein the incubation is carried out in the presence of a cell growth enhancing amount of protamine zinc insulin U.S.P., B.P. dispersed in said medium;
- removing the medium containing protamine zinc insulin after significant cell growth is obtained from the cells;
- replacing said medium with a tissue culture maintenance medium essentially free of protamine zinc insulin;
- prior to virus innoculation and continuing incubation of the cells in tissue culture.
A method which comprises inoculating a tissue culture growth medium with a suspension of living animal cells, said growth medium being an intimate mixture of:
- a. a chemically defined protein-free tissue culture maintenance medium which is adapted to maintain metabolism of said cells but which is insufficient to support growth of said cells into a substantially complete monolayer in tissue culture;
- and
- b. an amount of from about 0.05 to about 0.5 units per milliliter of protamine zinc insulin U.S.P., B.P. effective to provide an ultimate growth medium adapted to support such growth medium adapted to support such growth of said cells;
- and incubating said cells and said growth medium in tissue culture under conditions conducive to growth of a confluent cell monolayer and thereafter inoculating said cells with a virus.
In a method which comprises inoculating a tissue culture medium with a suspension of primary living animal cells and incubating the cells and medium in tissue culture until significant cell growth is obtained, prior to virus inoculation, the improvement wherein the medium is Medium 199 having a cell growth enhancing amount of protamine zinc insulin U.S.P., B.P. dispersed therein.
A chemcially defined tissue culture growth medium comprising Medium 199 having a cell growth enhancing amount of from about 0.05 to about 0.5 units of protamine zinc insulin U.S.P., B.P. per milliliter dispersed therein said zinc insulin providing essentially the sole source of detectable insulin activity therein, said medium containing from about 1 to about 2 grams glucose per liter.
Dependencies are inferred from the claim text.