MDCK suspension cell lines in serum-free, chemically-defined media for vaccine production
AssigneesNational Health Research Institutes, Fujifilm Biosciences Inc.
InventorsJenny Bang, Hsiao-Tzu Ni, Alan Yung-Chih Hu, Tsai-Chuan Weng
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Abstract
Disclosed is an adapted Madin-Darby canine kidney cell line capable of suspension culture in the absence of serum, and a chemically-defined medium for culture of the adapted MDCK cell line. Further disclosed are culture methods for growing the adapted MDCK cell line and methods for producing a vaccine from the adapted MDCK cell line grown in the chemically-defined medium.
Claims15
A method of culturing suspension-and serum-free-adapted Madin-Darby canine kidney (MDCK) cells comprising contacting suspension-and serum-free-adapted MDCK cells (i) without microcarriers;
- and (ii) with chemically-defined and animal-component-free-growth medium, wherein the contacting is between about 1 day and about 12 days without medium exchange, wherein the chemically-defined and animal-component-free-medium comprises about 20 mM to about 30 mM glucose, wherein the culturing is suspension culturing at 5% CO2.
A method for producing and culturing an isolated suspension of a serum free adapted Madin-Darby canine kidney (MDCK) cell line comprising:
- (a) contacting adherent MDCK cells with a growth medium comprising between about 1% v/v to about 20% v/v serum for a period of time sufficient to establish the adherent MDCK cells in culture;
- (b) contacting the established adherent MDCK cells with increasing amounts of chemically-defined growth medium and decreasing amounts of the serum serially until the chemically-defined growth medium does not comprise serum and represents all of the medium;
- (c) isolating the adapted MDCK cells thereby producing an isolated suspension of a serum free adapted MDCK cell line;
- and
- (d) culturing the isolated suspension of a serum free adapted MDCK cell line between about 1 day and about 12 days with a chemically-defined and animal-component-free-medium without medium exchange and without microcarriers.
Dependencies are inferred from the claim text.