A free-radical hypothesis for the instability and evolution of genotype and phenotypein vitro

Parchment, Ralph E.; Natarajan, Kunthavi

doi:10.1007/bf00570888

Cited by 11 publications

(5 citation statements)

References 212 publications

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“…Most anchorage-dependent cell lines had generation times of ≈12 h in the exponential growth phase. 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).…”

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

confidence: 80%

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-sfmentioning

confidence: 80%

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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“…Although they have high self-renewal capacity, dental stem cells such as SHED will ultimately enter an irreversible proliferation-arrested state referred to as replicative senescence after long term in vitro culture under culture stresses, such as hyperoxia and elevated temperature [ 6 ]. Several strategies have been explored to overcome this replicative senescence, such as treatment of SHED with benzopyrene (BaP), radiation, or ectopic expression of viral oncogenes [ 7 ].…”

Section: Introductionmentioning

confidence: 99%

A novel method for banking stem cells from human exfoliated deciduous teeth: lentiviral TERT immortalization and phenotypical analysis

Yin

Wang

et al. 2016

Stem Cell Res Ther

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BackgroundStem cells from human exfoliated deciduous teeth (SHED) have recently attracted attention as novel multipotential stem cell sources. However, their application is limited due to in vitro replicative senescence. Ectopic expression of telomerase reverse transcriptase (TERT) is a promising strategy for overcoming this replicative senescence. Nevertheless, its potential application and the phenotype as well as tumorigenicity have never been assessed in SHED.MethodsTERT expression was stably restored in SHED (TERT-SHED) isolated from healthy children aged 6–8 years using lentiviral transduction with a puromycin selection marker. The expression of TERT was detected using reverse transcription polymerase chain reaction, Western blot and immunofluorescence. Surface markers of SHED were detected by flow cytometry. Enzyme-linked immunosorbent assay was used to assess senescence-associated β-galactosidase, while CCK-8 methods were used to examine the proliferation capacity of SHED and TERT-SHED at different passages. Moreover, multilineage differentiation, karyotype, colony formation in soft agar, and tumor formation in nude mice of SHED and TERT-SHED were also examined.ResultsLentiviral transduction induced stable TERT expression even in SHED at the 40th passage. TERT-SHED showed robust proliferation capacity and low concentration of β-galactosidase. Although they had some different biomarkers than early passage SHED, TERT-SHED at late passage showed similar mutilineage differentiation as TERT at early passage. Moreover, TERT-SHED at late passage showed normal karyotype, no soft agar colony formation, and no tumor formation in nude mice.ConclusionsTERT-immortalized SHED may be a promising resource for stem-cell therapy, although attention should be paid to the biological behavior of the cells.

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“…However, several groups have recently provided indirect evidence that amine oxidases may be involved in tumorigenesis via production of reactive aldehydes and hydrogen peroxide [16,17]. Moreover, in earlier studies it has been demonstrated that hydrogen peroxide produced by amine oxidases may cause spontaneous neoplastic transformation of cultivated murine cells in vitro [3]. Notably, it seems that the specific activities of aldehyde dehydrogenases are reduced in benign prostate hyperplasia, as well as in prostate tumours [18].…”

Section: Discussionmentioning

confidence: 99%

“…Oxidation of these substrates yields formaldehyde and methylglyoxal respectively. In contrast with a protective role of SSAOs, it has been proposed by several investigators that reactive aldehydes and hydrogen peroxide produced by endogenous amine oxidases could be of importance with regard to carcinogenesis [3,4].…”

Section: Introductionmentioning

confidence: 97%

Elevated activity of semicarbazide-sensitive amine oxidase in blood from patients with skeletal metastases of prostate cancer

et al. 1999

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The semicarbazide-sensitive amine oxidases constitute a group of copper-containing enzymes whose physiological function is unclear. The enzymes are present in various tissues, including blood plasma. At present, the source of the plasma enzyme in humans is not known. Results of a recent study suggested that semicarbazide-sensitive amine oxidase is expressed in the skeleton, e.g. in the spine. Using an indirect autoradiographic method in mice, we provide evidence that semicarbazide-sensitive amine oxidase is present in high abundance in bone tissue. Specific activities of semicarbazide-sensitive amine oxidase were estimated in blood samples from subjects with femoral bone fractures. Moreover, enzyme activities were also measured in patients suffering from prostate cancer with skeletal metastases. The level of specific semicarbazide-sensitive amine oxidase activity in serum was significantly elevated in patients with skeletal metastases compared with both healthy controls and patients having prostate cancer without signs of skeletal metastases. Based on the results of the present study, we propose that semicarbazide-sensitive amine oxidase in blood plasma may originate, at least in part, from the skeleton.

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A free-radical hypothesis for the instability and evolution of genotype and phenotypein vitro

Cited by 11 publications

References 212 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

A novel method for banking stem cells from human exfoliated deciduous teeth: lentiviral TERT immortalization and phenotypical analysis

Elevated activity of semicarbazide-sensitive amine oxidase in blood from patients with skeletal metastases of prostate cancer

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