Temperature and medium composition were changed with the aim of increasing growth and erythropoietin (EPO) production in EPO-producing Chinese hamster ovary (CHO) cells. We used the CHO cell line, IBE, and its derivative, CO5, which over-expresses the first two enzymes of the urea cycle, carbamoyl p...
Temperature and medium composition were changed with the aim of increasing growth and erythropoietin (EPO) production in EPO-producing Chinese hamster ovary (CHO) cells. We used the CHO cell line, IBE, and its derivative, CO5, which over-expresses the first two enzymes of the urea cycle, carbamoyl phosphate synthetase I (CPS I) and ornithine transcar-bamoylase (OTC). When supplements were added to the medium at $33\;^{\circ}C$, the growth of IBE and CO5 cells increased by $27\%\;and;26\%$, respectively and the maximum yield of EPO was increased by $40\%$ in both cell lines. The absolute EPO concentration in the CO5 cells was always $55{\sim}60\%$ higher than in the IBE cells. In addition, when the two cell lines were continuously cultured with supplements at $33\;^{\circ}C$ until their growth rates approached those at $37\;^{\circ}C$, the growth rates of both IBE and CO5 cells increased by $54\%$ and their maximum EPO levels increased by up to $73\%\;and\;56\%$, respectively. Therefore, the growth and EPO expression levels of CO5 cells increased 2.2-fold and 2.6-fold, respectively, compared to those of the IBE cells. These results indicate that adaptation to lower temperature as well as medium supplementation could be important for improving cell growth and EPO production.
Temperature and medium composition were changed with the aim of increasing growth and erythropoietin (EPO) production in EPO-producing Chinese hamster ovary (CHO) cells. We used the CHO cell line, IBE, and its derivative, CO5, which over-expresses the first two enzymes of the urea cycle, carbamoyl phosphate synthetase I (CPS I) and ornithine transcar-bamoylase (OTC). When supplements were added to the medium at $33\;^{\circ}C$, the growth of IBE and CO5 cells increased by $27\%\;and;26\%$, respectively and the maximum yield of EPO was increased by $40\%$ in both cell lines. The absolute EPO concentration in the CO5 cells was always $55{\sim}60\%$ higher than in the IBE cells. In addition, when the two cell lines were continuously cultured with supplements at $33\;^{\circ}C$ until their growth rates approached those at $37\;^{\circ}C$, the growth rates of both IBE and CO5 cells increased by $54\%$ and their maximum EPO levels increased by up to $73\%\;and\;56\%$, respectively. Therefore, the growth and EPO expression levels of CO5 cells increased 2.2-fold and 2.6-fold, respectively, compared to those of the IBE cells. These results indicate that adaptation to lower temperature as well as medium supplementation could be important for improving cell growth and EPO production.
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이론/모형
The supernatants were aliquoted and standard solutions (Retaining 0, 1, 3 and 5 mM Nu4CI were prepared in the medium and stored at -20 ℃. Ammonia concentrations were determined by a modification of the previo너이y described indophenol method (Sikdar and Sawant, 1994). To remove inhibitors of the colorimetric reaction, proteinremoving reagents (1% sodium tungstate, 0.
성능/효과
In conclusion, cell viability and EPO expression levels increased in both CO5 and IBE cells cultured in the presence of serum-free medium plus supplements until their growth rate at 33 ℃ approached that at 37 ℃, suggesting that adaptation to the lower temperature as well as medium composition are important for improving cell growth and recombinant protein production. These results may be of wide applicable for increasing the production of other physiologically active proteins in mammalian cell cultures.
, and OTC cells. The results showed that the ammonia concentrations in the culture media (Fig. 3A, B, and C), as well as the concentrations per cell (data not shown), were not affected by temperature and medium composition and that those in CO5 and OTC cells, respectively, were 15-20% and 40% lower than in 旧E cells. This suggests that ammonia concentrations are essentially unaffected by temperature and medium composition.
2C). These results indicate that the growth of each of the cell lines cultured in the presence of serum-free medium plus supplements, and cultured at 33 ℃ until their growth rates were similar to those of cells at 37 ℃ increased by 20-30% and 55-60% respectively, compared to those of the cells grown in serum-free medium. The viability of CO5 and OTC cells was 기ways 20-30% and 40% higher than that of 旧E cells, respectively.
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