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Titolo:
Novel surface tagging technology for selection of complex proliferation-controlled mammalian cell phenotypes
Autore:
Schlatter, S; Bailey, JE; Fussenegger, M;
Indirizzi:
ETH Zurich, Inst Biotechnol, Swiss Fed Inst Technol, CH-8093 Zurich, Switzerland ETH Zurich Zurich Switzerland CH-8093 chnol, CH-8093 Zurich, Switzerland
Titolo Testata:
BIOTECHNOLOGY AND BIOENGINEERING
fascicolo: 5, volume: 75, anno: 2001,
pagine: 597 - 606
SICI:
0006-3592(200112)75:5<597:NSTTFS>2.0.ZU;2-7
Fonte:
ISI
Lingua:
ENG
Soggetto:
HAMSTER OVARY CELLS; GENE-EXPRESSION; CD34(+) CELLS; GAUCHER PATIENTS; BONE-MARROW; CHO CELLS; PRODUCTIVITY; GROWTH; APOPTOSIS; VECTORS;
Keywords:
CHO cells; proliferation control; surface selection; mammalian cell culture technology; SEAP; p27; hook; tetracycline; regulated expression; surface tagging; magnetic selection; multicistronic; FACS;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Agriculture,Biology & Environmental Sciences
Life Sciences
Citazioni:
50
Recensione:
Indirizzi per estratti:
Indirizzo: Fussenegger, M ETH Zurich, Inst Biotechnol, Swiss Fed Inst Technol, CH-8093 Zurich, Switzerland ETH Zurich Zurich Switzerland CH-8093 Zurich, Switzerland
Citazione:
S. Schlatter et al., "Novel surface tagging technology for selection of complex proliferation-controlled mammalian cell phenotypes", BIOTECH BIO, 75(5), 2001, pp. 597-606

Abstract

Regulated overexpression of the cyclin dependent kinase inhibitor p27 enables biphasic production processes which consist of a nonproducing expansionphase followed by an extended proliferation-arrested production phase. During the growth-arrested production phase proliferation-competent mutants emerge as a consequence of genetic drift and strong counterselection. Here, we evaluate the use of cell surface markers for ex vivo selection of growth-arrested phenotypes by magnetic or FACS-mediated cell sorting. Multigene metabolic engineering resulted in a Chinese hamster ovary- (CHO) derived cellline CHO-SS101(5), which expresses the model product protein SEAP (secreted alkaline phosphatase), the human cyclindependent kinase inhibitor p27, and a membrane-anchored multidomain surface marker Hook in a tricistronic tetracycline-repressible manner. In the absence of tetracycline in the cell culture medium, p27 mediated a Gl-phase-specific cell-cycle arrest of CHO-SS101(5) and resulted in a fivefold increase in SEAP production compared to proliferation-competent control cells. Concomitant expression of Hook enabledFACS- or magnetic-based selection of CHO-SS101(5) cells from various mixedpopulations. Surface selection of engineered cells will likely become important for biopharmaceutical manufacturing and for in vivo maintenance of treated cells in gene therapy and tissue engineering. (C) 2001 John Wiley & Sons, Inc.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 29/03/20 alle ore 08:44:47