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Titolo:
Serum protects protein-free competent Chinese hamster ovary cells against apoptosis induced by nutrient deprivation in batch culture
Autore:
Zanghi, JA; Fussenegger, M; Bailey, JE;
Indirizzi:
ETHch,nggerberg, Swiss Fed Inst Technol, Inst Biotechnol, HPT, CH-8093 Zuri ETH Honggerberg Zurich Switzerland CH-8093 Biotechnol, HPT, CH-8093 Zuri
Titolo Testata:
BIOTECHNOLOGY AND BIOENGINEERING
fascicolo: 1, volume: 64, anno: 1999,
pagine: 108 - 119
SICI:
0006-3592(19990705)64:1<108:SPPCCH>2.0.ZU;2-F
Fonte:
ISI
Lingua:
ENG
Soggetto:
GROWTH-FACTOR-I; MONOCLONAL-ANTIBODY PRODUCTION; HYBRIDOMA CELLS; AMINO-ACIDS; CHO CELLS; EXPRESSION; PROLIFERATION; BCL-2; SURVIVAL; LINE;
Keywords:
serum-free medium; protein-free medium; Chinese hamster ovary; apoptosis, cell death; cyclin E; cell-time; cell-hours;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Life Sciences
Citazioni:
41
Recensione:
Indirizzi per estratti:
Indirizzo: Bailey, JE ETHch,nggerberg, Swiss Fed Inst Technol, Inst Biotechnol, HPT, CH-8093 Zuri ETH Honggerberg Zurich Switzerland CH-8093 HPT, CH-8093 Zuri
Citazione:
J.A. Zanghi et al., "Serum protects protein-free competent Chinese hamster ovary cells against apoptosis induced by nutrient deprivation in batch culture", BIOTECH BIO, 64(1), 1999, pp. 108-119

Abstract

The development of serum- and protein-free Chinese hamster ovary (CHO) cell cultures is a high priority for the production of biopharmaceuticals. Protein-free competent CHO cells lines have been previously constructed by twodifferent methods-metabolic engineering with cell-cycle regulatory proteins and long-term selective adaptation. Apoptosis was present in both cell lines during protein-free, static-batch culture as a result of nutrient deprivation, and glucose deprivation alone was a potent inducer of apoptosis compared to the depletion of other nutrients such as amino acids. By adding back serum to the cultures during batch growth or nutrient deprivation, it was shown that unidentified survival factors in serum can greatly reduce apoptosis in protein-competent cell lines in all phases of the culture. Both observations contrast to previous reports for hybridoma cells, in which aminoacids were the key determinants of apoptosis and serum had no additional anti-apoptotic effect. Serum's protective effect against CHO cell death in batch culture was multifaceted and complex: (1) 10% FBS increased cell viability to >99% during exponential growth from roughly 75-90%, (2) 5-10% fetalbovine serum (FBS) reduced specific glucose consumption rates in both celllines by 40%, thereby delaying the onset of apoptosis caused by glucose deprivation, and (3) 5% FBS reduced the specific cell death rate by 65% during a 3-d lactate-consumption phase characterized by substantial abortive proliferation, in which the cells both proliferated and died at a constant rate. The benefit of serum on cell production over the various phases of batchgrowth was combined into a single parameter by integrating the viable cellconcentration vs, time profile (termed here as cumulative volumetric viable cell-time, VCTvol). Despite the ability of both cell lines to grow indefinitely without any exogenous growth factors, the addition of serum resultedin a 2.3-fold increase in the VCTvol. Thus, it is clear that there is muchroom for improvement of protein-free CHO cell lines despite their adequategrowth competence, and new strategies different from those successfully used for hybridomas may be necessary to combat CHO cell apoptosis. (C) 1999 John Wiley & Sons, Inc.

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Documento generato il 25/09/20 alle ore 06:25:20