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Titolo:
Regulated overexpression of the survival factor bcl-2 in CHO cells increases viable cell density in batch culture and decreases DNA release in extended fixed-bed cultivation
Autore:
Fussenegger, M; Fassnacht, D; Schwartz, R; Zanghi, JA; Graf, M; Bailey, JE; Portner, R;
Indirizzi:
ETH Zurich, Inst Biotechnol, CH-8093 Zurich, Switzerland ETH Zurich Zurich Switzerland CH-8093 chnol, CH-8093 Zurich, Switzerland Tech Univ Hamburg Harburg, D-21071 Hamburg, Germany Tech Univ Hamburg Harburg Hamburg Germany D-21071 21071 Hamburg, Germany
Titolo Testata:
CYTOTECHNOLOGY
fascicolo: 1, volume: 32, anno: 2000,
pagine: 45 - 61
SICI:
0920-9069(2000)32:1<45:ROOTSF>2.0.ZU;2-L
Fonte:
ISI
Lingua:
ENG
Soggetto:
HAMSTER OVARY CELLS; ENHANCED ANTIBODY-PRODUCTION; MAMMALIAN-CELLS; GENE-EXPRESSION; INDUCED APOPTOSIS; HYBRIDOMA CELLS; CYTOCHROME-C; MOLECULAR REGULATION; ANIMAL-CELLS; PROTEIN-FREE;
Keywords:
apoptosis; Bcl-2; fixed-bed reactor; regulated gene expression;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Citazioni:
72
Recensione:
Indirizzi per estratti:
Indirizzo: Bailey, JE ETH Zurich, Inst Biotechnol, CH-8093 Zurich, Switzerland ETH Zurich Zurich Switzerland CH-8093 093 Zurich, Switzerland
Citazione:
M. Fussenegger et al., "Regulated overexpression of the survival factor bcl-2 in CHO cells increases viable cell density in batch culture and decreases DNA release in extended fixed-bed cultivation", CYTOTECHNOL, 32(1), 2000, pp. 45-61

Abstract

Using multicistronic expression technology we generated a stable Chinese hamster ovary (CHO) cell line (MG12) expressing a model secreted heterologous glycoprotein, the secreted form of the human placental alkaline phosphatase (SEAP), and bcl-2, best known as an apoptosis inhibitor, in a tetracycline-repressible dicistronic configuration. In batch cultivations in serum-containing medium, MG12 cells reached twice the final viable cell density when Bcl-2 was overexpressed (in the absence of tetracycline) compared to MG12populations cultured under tetracycline-containing conditions (bcl-2 repressed). However, bcl-2-expressing MG12 cells showed no significant retardation of the decline phase compared to batch cultures in which the dicistronicexpression unit was repressed. Genetic linkage of bcl-2 expression with the reporter protein SEAP in our multicistronic construct allowed online monitoring of Bcl-2 expression over an extended, multistage fixed-bed bioreactor cultivation. The cloned multicistronic expression unit proved to be stable over a 100 day bioreactor run. CHO MG12 cells in the fixed-bed reactor showed a drastic decrease in the release of DNA into the culture supernatant under conditions of reduced tetracycline (and hence derepressed SEAP and bcl-2 overexpression). This observation indicated enhanced robustness associated with bcl-2 overexpression, similar to recent findings for constitutive Bcl-2-overexpressing hybridoma cells under the same bioprocess conditions. These findings indicate, in these serum-containing CHO cell cultures, that overexpression of Bcl-2 results in desirable modifications in culture physiology.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 23/09/20 alle ore 21:11:36