Effect of serum and sodium butyrate on erythropoietin production by recombinant Chinese hamster ovary cell = 재조합 CHO 세포에 의한 Erythropoietin의 생산에 미치는 혈청과 Sodium butyrate의 영향

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Effects of serum concentration on cellular responses of Chinese Hamster Ovary (CHO) cells were investigated to reduce serum concentration and produce EPO from lower serum media without a decrease of cell specific productivity. The cell specific productivity ($q_p$) and specific growth rate (μ) in 1.0% FBS were 4 and 5 fold lower than those in 10% FBS, respectively. Northern hybridization and MTT assay showed that transcription activity and cellular metabolic activity was proportional to serum concentration. The differences between 1.0% and 10% FBS culture in $\mu$ and $q_p$ were mainly due to the reduced cellular metabolism and mRNA synthesis activity. The relationship between cell cycle and specific productivity was unlike hybridoma which had the highest productivity in $G_0/G_1$ phase. On the contrary, the $G_0/G_1$ phase in CHO cell was the least productive phase. When the cells adapted in 10\% FBS were transferred to 1% FBS and cultured for 15 days by daily medium exchange, the specific productivity was constantly maintained over 15 days, regardless of a decrease of cellular metabolic activity. For the long term production of EPO with lower serum media, CHO cells were cultured in spinner flasks containing each of two types of microcarrier, Cytodex (3g/L) and CutispherG (1g/L). Media were replaced by 0.7 v/v/d, reducing stepwise serum concentration from 10% to 0.1% FBS over 50 days. Viable cell density did not decrease regardless of low serum concentration. EPO production was dependent on serum concentration. EPO concentration in 0.1% FBS was 4 fold lower than that in 10% FBS. Cells on Cytodex3 detached and formed cell aggregate according to culture time. However, entrapped in CultisoerG did not form cell aggregates Effect of sodium butyrate on EPO production was studied. Sodium butyrate, at millimolar concentration, when added to the culture medium in the inoculation period, arrested cell growth, but enhanced the specific productivity of EPO up to 12-...
Kim, Jung-Hoeresearcher김정회researcher
한국과학기술원 : 생물과학과,
Issue Date
114639/325007 / 000925030

학위논문(박사) - 한국과학기술원 : 생물과학과, 1997.2, [ vii, 116 p. ]


Serum; Erythropoietin; CHO cell; Sodium butyrate; 비생산성; 적혈구생성 호르몬; 혈청; 초세포; Specific productivity

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