Stability and cytogenetic characterization of recombinant CHO cell lines established by microinjection and calcium phosphate transfection

被引：2

作者：

Derouazi, M. ^{[1
]}

Martinet, D. ^{[2
]}

Besuchet, N. ^{[2
]}

Flaction, R. ^{[1
]}

Wicht, M. ^{[2
]}

Bertschinger, M. ^{[1
]}

Hacker, D. ^{[1
]}

Beckmann, J. ^{[2
]}

Wurm, F. M. ^{[1
]}

机构：

[1] Ecole Polytech Fed Lausanne, Fac Life Sci, Lab Cellular Biotechnol, CH-1015 Lausanne, Switzerland

[2] CHU Vaudois, Lab Cytogenet Serv Genet Med, CH-1011 Lausanne, Switzerland

来源：

CELL TECHNOLOGY FOR CELL PRODUCTS | 2007年

关键词：

CHO DG44; karyotype; recombinant proteins; calcium phosphate; microinjection; GFP;

D O I：

10.1007/978-1-4020-5476-1_73

中图分类号：

Q81 [生物工程学（生物技术）]; Q93 [微生物学];

学科分类号：

071005 ; 0836 ; 090102 ; 100705 ;

摘要：

Chinese hamster ovary cells (CHO) are widely used for the stable production of recombinant proteins. Typically, recombinant cell lines are characterized for the stability of protein expression over a period corresponding to the time needed to scale-up the culture and harvest the product (e.g. 2 to 3 months), for the number of plasmid copies integrated into the host genome, and for the quality and quantity of the recombinant protein. In this study we extended the characterization to the cytogenetic level. Sixteen recombinant CHO cell lines were established using calcium phosphate transfection and microinjection as DNA transfer methods. For each cell line we observed by fluorescence in situ hybridization a single integration site regardless of the gene delivery method, the topology of the DNA (circular or linear), or the integrated plasmid copy number (between 1 and 50). Integration was not targeted to a specific chromosome. Chromosomal rearrangements were observed in about half of these cell lines. This phenomenon occurred independently of the gene transfer method. Interestingly the rearrangements were not on the chromosome where the plasmid integrated. We observed rearrangements between chromosomes and chromosomal imbalances.

引用

页码：443 / +

页数：2