Literature DB >> 25087075

Combination of dextran sulfate and recombinant trypsin on aggregation of Chinese hamster ovary cells.

Yu Jing1,2,3,4, Cunchao Zhang1,2,3, Tuo Fu1,2,3, Cheng Jiang1,2,3, Kai Ma1,2,3, Dapeng Zhang1,2,3, Sheng Hou1,2,3, Jianxin Dai1,2,3, Hao Wang1,2,3, Xueguang Zhang4, Geng Kou5,6,7, Yajun Guo8,9,10,11.   

Abstract

In laboratory scale therapeutical protein production, cell clumps form typically in shake flasks, which hinders cell growth and decreases protein yield. To minimize clumps during the culture of Chinese hamster ovary cells, we employed the combination of two reagents, dextran sulfate (DS) and recombinant trypsin (r-trypsin). Our results showed that both DS and r-trypsin could diminish cell aggregation when adding them respectively, but clumps were still noticed obviously. In order to further mitigate cell agglomerate, a combination of 1.2 g/L DS and 8.0 mg/L r-trypsin was employed and no clumps were found under the bright field microscope. Strikingly, the highest viable cell density of combination group was increased from 5.12 × 10(6) to 7.13 × 10(6) cells/mL, while the integral of viable cells concentration was raised from 35.13 × 10(6) to 60.87 × 10(6) cells·days/mL, and the culture period was prolonged by 4 days. In addition, the antibody integrity was maintained in the combination group compared with that of the control.

Entities:  

Keywords:  Aggregation; Chinese hamster ovary cells; Dextran sulfate; Fed-batch; Recombinant trypsin

Year:  2014        PMID: 25087075      PMCID: PMC4754253          DOI: 10.1007/s10616-014-9774-4

Source DB:  PubMed          Journal:  Cytotechnology        ISSN: 0920-9069            Impact factor:   2.058


  12 in total

1.  Development of a shaking bioreactor system for animal cell cultures.

Authors:  C -M. Liu; L -N. Hong
Journal:  Biochem Eng J       Date:  2001-03       Impact factor: 3.978

2.  The growth factor inhibitor suramin reduces apoptosis and cell aggregation in protein-free CHO cell batch cultures.

Authors:  J A Zanghi; W A Renner; J E Bailey; M Fussenegger
Journal:  Biotechnol Prog       Date:  2000 May-Jun

Review 3.  Production of recombinant protein therapeutics in cultivated mammalian cells.

Authors:  Florian M Wurm
Journal:  Nat Biotechnol       Date:  2004-11       Impact factor: 54.908

4.  Expression of anti-apoptosis genes alters lactate metabolism of Chinese Hamster Ovary cells in culture.

Authors:  Haimanti Dorai; Yun Seung Kyung; Dawn Ellis; CherylAnn Kinney; Chengbin Lin; David Jan; Gordon Moore; Michael J Betenbaugh
Journal:  Biotechnol Bioeng       Date:  2009-06-15       Impact factor: 4.530

5.  Isolation and culture of primary equine tracheal epithelial cells.

Authors:  Workineh Shibeshi; Getu Abraham; Carsten Kneuer; Christin Ellenberger; Johannes Seeger; Heinz-Adolf Schoon; Fritz R Ungemach
Journal:  In Vitro Cell Dev Biol Anim       Date:  2008-07-02       Impact factor: 2.416

6.  Gene expression profiling for mechanistic understanding of cellular aggregation in mammalian cell perfusion cultures.

Authors:  Meile Liu; Chetan T Goudar
Journal:  Biotechnol Bioeng       Date:  2012-10-18       Impact factor: 4.530

7.  On the recovery of adhesiveness by trypsin-dissociated cells.

Authors:  M S Steinberg; P B Armstrong; R E Granger
Journal:  J Membr Biol       Date:  1973       Impact factor: 1.843

8.  Heparin promotes suspension adaptation process of CHO-TS28 cells by eliminating cell aggregation.

Authors:  Ling Li; Jun Qin; Qiang Feng; Hao Tang; Rong Liu; Liqing Xu; Zhinan Chen
Journal:  Mol Biotechnol       Date:  2011-01       Impact factor: 2.695

9.  Addition of exogenous protease facilitates reovirus infection in many restrictive cells.

Authors:  Joseph W Golden; Jessica Linke; Stephen Schmechel; Kara Thoemke; Leslie A Schiff
Journal:  J Virol       Date:  2002-08       Impact factor: 5.103

10.  Trypsinized BHK21 cells aggregate in the presence of metabolic inhibitors and in the absence of divalent cations.

Authors:  J G Edwards; J A Campbell; R T Robson; M G Vicker
Journal:  J Cell Sci       Date:  1975-12       Impact factor: 5.285

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