Abstract

A chemically defined protein free medium, DF6S, was developed for the cultivation of a recombinant Chinese hamster ovary cell line (CHO2DS) producing human prothrombin in suspension batch culture. DF6S was formulated by optimizing DME/F12 with amino acids and supplementing the optimized DME/F12 with aurintricarboxylic acid, ethanolamine, ferric sulfate, Pluronic F68, putrescine and sodium pyruvate. From a seeding density of 2.3 × 105 cells ml−1, CHO2DS cells grown in suspension in DF6S medium reached a maximal cell density of 1.92 × 106 cells ml−1 with an accumulated prothrombin concentration of 16.7 mg l−1 after 6 days in culture.

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