Development of a serum-free medium for the production of erythropoietin by suspension culture of recombinant Chinese hamster ovary cells using a statistical design

Lee, Gyun Min; Kim, Eun Jung; Kim, No Soo; Yoon, Seong Jun; Ahn, Yeon Soon; Song, Ji Yong

doi:10.1016/s0168-1656(99)00004-8

Cited by 62 publications

(43 citation statements)

References 20 publications

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“…Indeed, serum contains many components (growth factors, insulin, transferrin, albumin, vitamins, etc....), which are known to be essential to several cellular functions like growth regulation or intracellular transports (Froud 1999). Thus, compared to serum-containing cultures, reduction of protein production and/or cell growth are generally observed when cells are transferred to serum-free media (Inoue et al 1996;Lee et al 1999;Ozturk et al 2003). Extensive investigations for optimising medium composition to a determined cell line cultured in the desired conditions are required to recover equivalent performances (Buntemeyer et al 1991;Hewlett 1991;Castro et al 1992;Liu et al 2001).…”

Section: Introductionmentioning

confidence: 99%

Related effects of cell adaptation to serum-free conditions on murine EPO production and glycosylation by CHO cells

et al. 2006

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The necessity to perform serum-free cultures to produce recombinant glycoproteins generally requires an adaptation procedure of the cell line to new environmental conditions, which may therefore induce quantitative and qualitative effects on the product, particularly on its glycosylation. In previous studies, desialylation of EPO produced by CHO cells was shown to be dependent on the presence of serum in the medium. In this paper, to discriminate between the effects of the adaptation procedure to serum-free medium and the effects of the absence of serum on EPO production and glycosylation, adapted and nonadapted CHO cells were grown in serum-free and serum-containing media. The main kinetics of CHO cells were determined over batch processes as well as the glycosylation patterns of produced EPO by HPCE-LIF. A reversible decrease in EPO production was observed when cells were adapted to SFX-CHO TM medium, as the same cells partially recovered their production capacity when cultivated in serum-containing medium or in the enriched SFM TM serum-free medium. More interestingly, EPO desialylation that was not observed in both serum-free media was restored if the serum-independent cells were recultured in presence of serum. In the same way, while the serum-independent cells did not release a sialidase activity in both serum-free media, a significant activity was recovered when serum was added. In fact, the cell adaptation process to serum-free conditions did not specifically affect the sialidase release and the cellular mechanism of protein desialylation, which appeared to be mainly related to the presence of serum for both adapted and non-adapted cells.

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Section: Introductionmentioning

confidence: 99%

Related effects of cell adaptation to serum-free conditions on murine EPO production and glycosylation by CHO cells

et al. 2006

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“…However, the serum-free medium formulation used in this study was not published. In addition, many formulations for recombinant CHO cells expressing a variety of proteins are described (Asakura et al, 1992;Blüml et al, 1994;Hata et al, 1992;Ii et al, 1995;Kim et al, 1998;Kitano, 1994;Lee et al, 1999;Matthews et al, 1994;Mizuguchi et al, 1993;Scharfenberg and Wagner, 1995;Schorn et al, 1994;Wyatt, 1994). However, a versatile serum-free medium formulation for the parental cell line DUKXB11 has not been reported to date.…”

Section: Introductionmentioning

confidence: 99%

Serum- and protein-free media formulations for the Chinese hamster ovary cell line DUKXB11

Schröder

Matischak

Friedl

2004

Journal of Biotechnology

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Citation for published item:hr¤ oderD wF nd w tis h kD uF nd priedlD F @PHHRA 9 erumE nd proteinEfree medi formul tions for the ghinese h mster ov ry ell line h u fIIF9D tourn l of iote hnologyFD IHV @QAF ppF PUWEPWPF Further information on publisher's website: httpXGGdxFdoiForgGIHFIHITGjFj iote FPHHQFIPFHHS Publisher's copyright statement:Additional information: Use policyThe full-text may be used and/or reproduced, and given to third parties in any format or medium, without prior permission or charge, for personal research or study, educational, or not-for-pro t purposes provided that:• a full bibliographic reference is made to the original source • a link is made to the metadata record in DRO • the full-text is not changed in any way The full-text must not be sold in any format or medium without the formal permission of the copyright holders.Please consult the full DRO policy for further details. AbstractThe production of therapeutic proteins in mammalian cell lines is of outstanding importance. The maintenance of most mammalian cell lines in culture requires the addition of serum to the culture medium.The elimination of serum from mammalian cell culture is desirable since serum is expensive and a source of contaminants, e.g. viruses, mycoplasma or prions. Here we describe the composition of serum-and protein-free media for the Chinese hamster ovary (CHO) cell line DUKXB11. The serum-free formulation supports excellent growth of CHO DUKXB11 cells at low (23 cells/cm 2 ) and high (2·10 4 cells/cm 2 ) seeding densities characterized by a generation time of 10 -12 h, and, after addition of 0.2% pluronic F-68, the growth of a recombinant suspension cell line derived from DUKXB11. In addition, this formulation also allowed us to adapt recombinant cell lines expressing various amounts of human antithrombin ATIII (ATIII)to serum-free conditions. Secretion of ATIII was readily observed in the serum-free medium. Minor changes to the serum-free formulation resulted in a protein free formulation that supported growth of CHO DUKXB11 cells, growth of recombinant CHO cells expressing ATIII, and production of ATIII. Key wordsChinese hamster ovary cells, serum-free medium, protein-free medium, recombinant protein production,

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“…Classical experimental design requires only one growth factor being changed in the growth medium at a time to determine its contribution in cellular activity. In the statistical screening protocol, the widely accepted medium optimization tool, Plackett-Burman design (Plackett and Burman 1946) is used as multifactorial statistical design (Stanbury et al 1986) that efficiently screens the important factors among a large number of variables and accounts for the interactions between them (Castro et al 1992;Lee et al 1999;González-Leal et al 2011;Zhang et al 2013). However, Plackett-Burman design for screening growth factors for enhanced growth and proliferation of shrimp cells in vitro has not been adapted.…”

Section: Introductionmentioning

confidence: 99%

Multifactorial interaction of growth factors on Penaeus monodon lymphoid cells and the impact of IGFs in DNA synthesis and metabolic activity in vitro

2014

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Development of continuous cell lines from shrimp is essential to investigate viral pathogens. Unfortunately, there is no valid cell line developed from crustaceans in general and shrimps in particular to address this issue. Lack of information on the requirements of cells in vitro limits the success of developing a cell line, where the microenvironment of a cell culture, provided by the growth medium, is of prime importance. Screening and optimization of growth medium components based on statistical experimental designs have been widely used for improving the efficacy of cell culture media. Accordingly, we applied Plackett-Burman design and response surface methodology to study multifactorial interactions between the growth factors in shrimp cell culture medium and to identify the most important ones for growth of lymphoid cell culture from Penaeus monodon. The statistical screening and optimization indicated that insulin like growth factor-I (IGF-I) and insulin like growth factor-II (IGF-II) at concentrations of 100 and 150 ng ml -1 , respectively, could significantly influence the metabolic activity and DNA synthesis of the lymphoid cells. An increase of 53 % metabolic activity and 24.8 % DNA synthesis could be obtained, which suggested that IGF-I and IGF-II had critical roles in metabolic activity and DNA synthesis of shrimp lymphoid cells.

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Development of a serum-free medium for the production of erythropoietin by suspension culture of recombinant Chinese hamster ovary cells using a statistical design

Cited by 62 publications

References 20 publications

Related effects of cell adaptation to serum-free conditions on murine EPO production and glycosylation by CHO cells

Related effects of cell adaptation to serum-free conditions on murine EPO production and glycosylation by CHO cells

Serum- and protein-free media formulations for the Chinese hamster ovary cell line DUKXB11

Multifactorial interaction of growth factors on Penaeus monodon lymphoid cells and the impact of IGFs in DNA synthesis and metabolic activity in vitro

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