Osmoprotective effect of glycine betaine on thrombopoietin production in hyperosmotic Chinese hamster ovary cell culture: Clonal variations

When 23 recombinant Chinese hamster ovary (rCHO) cell clones were cultivate d in hyperosmolar medium resulting from NaCl addition (533 mOsm/kg), their specific thrombopoietin (TPO) productivity (q(TPO)) was increased. However, due to depressed cell growth at elevated osmolality, no enhancement in the maximum TPO titer was made in batch cultures of all 23 clones. To test the feasibility of using glycine betaine, known as a strong osmoprotective com pound, for improved TPO production in hyperosmotic rCHO cell cultures, hype rosmotic batch cultures of 23 clones were performed in the presence of 15 m M glycine betaine. Glycine betaine was found to have a strong osmoprotectiv e effect on all 23 clones. Inclusion of 15 mM glycine betaine in hyperosmol ar medium enabled 22 clones to grow at 542 mOsm/kg, where most clones could not grow in the absence of glycine betaine, but at a cost of reduced qTPO However, the relative decrease in q(TPO) varied significantly among clones. Thus, efficacy of the simultaneous use of hyperosmotic pressure and glycin e betaine as a means to improve foreign protein production was variable amo ng clones. Six out of 23 clones displayed more than a 40% increase in the m aximum TPO titer in the hyperosmolar medium containing glycine betaine, com pared with that in the standard medium with a physiological osmolality. Tak en together, the results obtained here emphasize the importance of selectio n of clones for the successful use of hyperosmotic pressure and glycine bet aine as an economical means to improve TPO production.