Lactate dehydrogenase (LDH) activity of the number of dead cells in the medium of cultured eukaryotic cells as marker

Legrand, Chantal; Jm, Bour; Jacob, Claus; Capiaumont, J.; Martial, A.; Marc, Annie; Wudtke, M.; Kretzmer, G.; Demangel, Caroline; Duval, Dominique

doi:10.1016/0168-1656(92)90158-6

Cited by 357 publications

(157 citation statements)

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“…This prompted us to investigate the degree of cell lysis in medium CHO-T1-SF after inoculation at higher cell densities (2·10 4 cells/cm 2 ). The extent of cell lysis during the cultivation of mammalian cells can be determined by measuring the activity of L-LDH in conditioned media (Bour et al, 1988;Goergen et al, Schröder, Matischak, and Friedl 14 1993;Jauregul et al, 1981;Legrand et al, 1992;Racher et al, 1990). However, we were not able to detect any L-LDH activity in media conditioned by high-density cultures of any one of the anchorage- (Schröder and Friedl, 1997a).…”

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

confidence: 43%

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-sfcontrasting

confidence: 43%

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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“…Cells were co-transfected with pAP-tag4 (GenHunter, Nashville, TN) as a control for transfection efficiency. After 48 h, cell death was analyzed by measuring lactate dehydrogenase (LDH) in the cell medium using a LDH cytotoxicity detection kit (Takara) according to the manufacturer's protocol (Decker and Lohmann-Matthes, 1988;Legrand et al, 1992). Percent cell death was calculated as 100% x [(LDH activity for the test condition -LDH activity for the negative control)/LDH activity in cells treated with 1% Triton X-100 (positive control)].…”

Section: Induction Of Apoptosis By Fasl Fusion Proteinsmentioning

confidence: 99%

Improvement of FasL Gene Therapy In Vitro by Fusing the FasL to Del1 Protein Domains

Kitano¹,

Mamiya²,

Hidai³

2011

Targets in Gene Therapy

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“…The presence of LDH in culture media can be used to predict the physiological state of different types of cell lines. It is known that there is a quantitative correlation between LDH leakage and the number of dead cells; therefore, an increase in LDH leakage can be used as a marker of cytotoxicity (Legrand et al, 1992).…”

Section: Introductionmentioning

confidence: 99%

Comparison of phototoxic effects of hypericin-mediated photodynamictherapy in HT-29 and Caco-2 colon cancer cells

Süloğlu¹,

Selmanoğlu²,

Yılmaz³

et al. 2016

Turk J Biol

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Hypericin (HYP) is a plant-derived photosensitizer. HYP is preferentially taken up by tumor cells. We designed this study to compare HYP-mediated photodynamic therapy (PDT) in HT-29 and Caco-2 colon cell lines. Cells were treated with 0.04, 0.08, and 0.15 µM HYP concentrations and irradiated. The effect of HYP on metabolic profiles, alterations in lactate dehydrogenase (LDH) leakage, and cell cycle progression was investigated for the first time. Changes in glucose consumption, lactate production, and LDH leakage were analyzed. HYP-induced cell death was quantified by double staining (acridine orange/propidium iodide) and alterations in cell cycle regulation were analyzed with flow cytometry (using propidium iodide). LDH leakage and the number of dead cells were elevated, and glucose consumption and lactate production decreased in a dose-and time-dependent manner. PDT resulted in an induction of apoptosis, mostly at the 0.08 µM HYP concentration. Apoptosis and/or necrosis were increased in the 0.15 µM HYP group. The accumulation of cells in the G2/M phase might account for the growth inhibition in HT-29 and Caco-2 cells with 0.08 µM HYP photoactivation. The observed G2/M arrest suggested that HYP may slow down the growth of colon cancer cells by regulating the cell cycle, leading either to growth inhibition or to initiation of apoptotic pathways.

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Lactate dehydrogenase (LDH) activity of the number of dead cells in the medium of cultured eukaryotic cells as marker

Cited by 357 publications

References 20 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

Improvement of FasL Gene Therapy In Vitro by Fusing the FasL to Del1 Protein Domains

Comparison of phototoxic effects of hypericin-mediated photodynamictherapy in HT-29 and Caco-2 colon cancer cells

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