Long-term multiplication of the Chinese hamster ovary (CHO) cell line in a serum-free medium

Gasser, F.; Mulsant, Philippe; Gillois, M.

doi:10.1007/bf02620890

Cited by 39 publications

(16 citation statements)

References 23 publications

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“…1 samples for cell counting were withdrawn. We observed a generation time of ≈25 h in CHO-T1-SF medium, a value that is comparable to other results published for CHO cells grown in suspension (Gasser et al, 1985;Lee et al, 1996;Renner et al, 1995;Sharif and Stevenson, 1993;Sinacore et al, 1996).…”

Section: Growth Characteristics In Serum-free Medium Cho-t1-sfsupporting

confidence: 91%

“…These values show no statistically significant differences to the colony forming efficiency seen in the presence of 10% (v/v) FCS (Gasser et al, 1985). When conditioned CHO-T1-SF medium plus 0.1% (w/v) ExCyte III was used, slightly lower colony forming efficiencies were observed than with fresh CHO-T1-SF medium (Table 2).…”

Section: Colony Forming Efficiencies In Medium Cho-t1-sfmentioning

confidence: 80%

“…This is the optimum for rodent cells in vitro (Parchment and Natarajan, 1992). For other serum-free media generation times of ≈24 h were reported (Gasser et al, 1985;Lee et al, 1996;Renner et al, 1995;Sharif and Stevenson, 1993;Sinacore et al, 1996). Only for medium UC203 generation times similar to those observed in medium CHO-T1-SF were reported (Ii et al, 1995).…”

Section: Growth Characteristics In Serum-free Medium Cho-t1-sfmentioning

confidence: 95%

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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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Section: Growth Characteristics In Serum-free Medium Cho-t1-sfsupporting

confidence: 91%

Section: Colony Forming Efficiencies In Medium Cho-t1-sfmentioning

confidence: 80%

Section: Growth Characteristics In Serum-free Medium Cho-t1-sfmentioning

confidence: 95%

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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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“…LF (R5 mm) were recovered 96 h after the last altrenogest feeding. Granulosa cells were collected from individual follicle in MEM121/F12 (v/v) medium (Gasser et al 1985). A small aliquot fraction was examined using Feulgen colouration to select cells only from healthy follicles (frequent mitosis, no pyknosis), as described previously (Monget et al 1993, Besnard et al 1996.…”

Section: Granulosa Cell Isolation and Rna Extractionmentioning

confidence: 99%

In vivo gene expression in granulosa cells during pig terminal follicular development

Bonnet¹,

Cao²,

SanCristobal³

et al. 2008

Reproduction

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Ovarian antral follicular development is clearly dependent on pituitary gonadotrophins FSH and LH. Although the endocrine mechanism that controls ovarian folliculogenesis leading to ovulation is quite well understood, the detailed mechanisms and molecular determinants in the different follicular compartments remain to be clarified. The aim of this study was to identify the genes differentially expressed in pig granulosa cells along the terminal ovarian follicle growth, to gain a comprehensive view of these molecular mechanisms. First, we developed a specific micro-array using cDNAs from suppression subtractive hybridization libraries (345 contigs) obtained by comparison of three follicle size classes: small, medium and large antral healthy follicles. In a second step, a transcriptomic analysis using cDNA probes from these three follicle classes identified 79 differentially expressed transcripts along the terminal follicular growth and 26 predictive genes of size classes. The differential expression of 18 genes has been controlled using real-time PCR experiments validating the micro-array analysis. Finally, the integration of the data using Ingenuity Pathways Analysis identified five gene networks providing descriptive elements of the terminal follicular development. Specifically, we observed: (1) the down-expression of ribosomal protein genes, (2) the genes involved in lipid metabolism and (3) the down-expression of cell morphology and ion-binding genes. In conclusion, this study gives new insight into the gene expression during pig terminal follicular growth in vivo and suggested, in particular, a morphological change in pig granulosa cells accompanying terminal follicular growth. Reproduction (2008) 136 211-224

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“…These stages correspond to the early and mid-follicular phase. Ovaries were dissected, follicles were extracted and granulosa cells were recovered as individual samples, in 200 ml of MEM121/F12 (v/v) medium (Gasser et al, 1985). For each sample, 5 ml of the granulosa-cell suspension was smeared onto histological slides, fixed for 10 min in methanol-formaldehyde-acetic acid (80 : 15 : 5), and subsequently stained with Feulgen, as previously described (Besnard et al, 1996).…”

Section: Datamentioning

confidence: 99%

A pilot study on transcriptome data analysis of folliculogenesis in pigs

Tosser‐Klopp

Cao

Bonnet

et al. 2009

Animal

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Three different stages of pig antral follicles have been studied in a granulosa-cell transcriptome analysis on nylon microarrays (1152 clones). The data have been generated from seven RNA follicle pools and several technical replicates were made. The objective of this paper was to state the feasibility of a transcriptomic protocol for the study of folliculogenesis in the pig. A statistical analysis was chosen, relying on the linear mixed model (LMM) paradigm. Low variability within technical replicates was hence checked with a LMM. Relevant genes that might be involved in the studied process were then selected. For the most significant genes, statistical methods such as principal component analysis and unsupervised hierarchical clustering were applied to assess their relevance, and a random forest analysis proved their predictive value. The selection of genes was consistent with previous studies and also allowed the identification of new genes whose role in pig folliculogenesis will be further investigated.

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Long-term multiplication of the Chinese hamster ovary (CHO) cell line in a serum-free medium

Cited by 39 publications

References 23 publications

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

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

In vivo gene expression in granulosa cells during pig terminal follicular development

A pilot study on transcriptome data analysis of folliculogenesis in pigs

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