Viability measurements of hybridoma cells in suspension cultures

Coco-Martin, J.M.; Oberink, Jan W.; Groot, Tiny A. M. van der Velden-de; Beuvery, E. C.

doi:10.1007/bf02540030

Cited by 24 publications

(16 citation statements)

References 20 publications

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“…The optical alignment was checked each day, and all other settings were unchanged throughout the study. The viable and dead cells were separated on the basis of the fonvard light scatter to estimate the cell size, and the side angle light scatter (90 ES) was used to estimate the granulometry of the cells (8). Cell viability was also evaluated by the DNA content after ethidium bromide labeling (20).…”

Section: Co)mentioning

confidence: 99%

Effects of H2O2 on the growth, secretion, and metabolism of hybridoma cells in culture

Ostrovidov¹,

Franck²,

Capiaumont³

et al. 1998

In Vitro Cell.Dev.Biol.-Animal

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The effect of low concentrations of hydrogen peroxide (H2O2) (5 x 10(-7)-9.5 x 10(-7) M) on cell growth and antibody production was investigated with murine hybridoma cells (Mark 3 and anti-hPL) in culture. Cell growth, measured by flow cytometry with morphological parameters, was significantly stimulated by H2O2 (8 x 10(-7) M) but H2O2 concentration of 7 x 10(-6) M and above increased cell death. H2O2 stimulation of antibody production was nonsignificant. The metabolism of cells treated with 8 x 10(-7) or 1 x 10(-5) M H2O2 was similar to that of the control in terms of glucose and glutamine consumption, lactate and ammonia production, and amino acid concentrations in the medium. The concentrations of lactate dehydrogenase, a marker of cell death, in test and control cells were similar. However, concentrations of intracellular free radicals measured by flow cytometry with dihydrorhodamine 123 (DHR 123) and dichlorofluorescein diacetate (DCFH-DA) as fluorochromes were different. The reactive oxygen species content of cells in 8 x 10(-7) M H2O2 was similar to that of the controls, but there was a sudden, marked production of superoxide anions (detected with DHR 123) and H2O2 or peroxides (detected with DCFH-DA) by cells incubated with 1 x 10(-5) M H2O2 which increased with increasing H2O2 until cell death.

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Section: Co)mentioning

confidence: 99%

Effects of H2O2 on the growth, secretion, and metabolism of hybridoma cells in culture

Ostrovidov¹,

Franck²,

Capiaumont³

et al. 1998

In Vitro Cell.Dev.Biol.-Animal

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“…PI is a standard tool to assess cell viability in cultured cells (Bevensee et al 1995;Coco-Martin et al 1992;Juurlink and Hertz 1993;Loo and Rillema 1998). It has also been used as a cell death marker in some recent studies on cultured brain slices (Laake et al 1999;Lahtinen et al 2001;Sakaguchi et al 1997;Zimmer et al 2000), but rather rarely in situ (Inglefield and Schwartz-Bloom 1998;Scarabelli et al 1999;Tekkök and Goldberg 2001;Wolff et al 2000).…”

Section: Introductionmentioning

confidence: 99%

Dynamic Recording of Cell Death in the In Vitro Dorsal Vagal Nucleus of Rats in Response to Metabolic Arrest

Müller

Ballanyi

2003

Journal of Neurophysiology

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Müller, Michael and Klaus Ballanyi. Dynamic recording of cell death in the in vitro dorsal vagal nucleus of rats in response to metabolic arrest. J Neurophysiol 89: 551-561, 2003. 10.1152/jn.00559.2002. Anoxic/ischemic neuronal death is usually assessed in cell cultures or in vivo within a time window of 24 h to several days using the nucleic acid stain propidium iodide or histological techniques. Accordingly, there is limited information on the time course of such neuronal death. We loaded acute rat brain stem slices with propidium iodide for dynamic fluorometric recording of metabolic arrest-related cell death in the dorsal vagal nucleus. This model was chosen because dorsal vagal neurons show a graded response to metabolic inhibition: anoxia and aglycemia cause a sustained hyperpolarization, whereas ischemia induces a glutamate-mediated, irreversible depolarization. We found that the number of propidium iodide-labeled cells increased from 27% to 43% of total cell count within 1-7 h after preparation of slices. Compared with these untreated control slices, cyanide-induced anoxia (30 min) or aglycemia (1 h) did not cause further cell death, whereas 3-h aglycemia destroyed an additional 13% of cells. Ischemia (1 h) due to cyanide plus iodoacetate immediately labeled an additional 20% of cells, and an additional 48% of cells were destroyed within the following 3 h of postischemia. Continuous recording of propidium iodide fluorescence showed that loss of membrane integrity started within 25 min after onset of the ischemic depolarization and the concomitant intracellular Ca 2ϩ rise. The results show that propidium iodide can be used to monitor cell death in acute brain slices. Our findings suggest that pronounced cell death occurs within a period of 1-4 h after onset of metabolic arrest and is apparently due to necrotic/oncotic mechanisms.

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“…It was mixed with the cells 1:1. In this method, viable and non-viable cells were counted on hemocytometer (7).…”

Section: Application Of Solutionsmentioning

confidence: 99%

Dextrose solution used for prolotherapy decreases cell viability and increases gene expressions of angiogenic and apopitotic factors

Guran¹,

Çoban²,

Karasimav³

et al. 2018

Gulhane Med J

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Dextrose solutions (in 1%, 5%, 10%, 15%, 20% and 25% concentrations) were applied on adult human fibroblasts in vitro culture condition. Cell proliferation assay was used for finding the cell deaths in cell culture. Up to 80% fibroblast cells died in high dextrose concentrations (15%, 20% and 25%). The deaths of cells were visualized on inverted microscope. In low dextrose concentrations (1%, 5% and 10%), viable cell ratio was above 80%. The gene expression analysis were performed on selected genes which have roles on angiogenesis and apopitosis [VEGFA (Vascular Endothelial Growth Factor A-OMIM ABSTRACTObjectives: Hypertonic dextrose injection in prolotherapy is an injection-based treatment used in chronic musculoskeletal conditions. Dextrose prolotherapy raises growth factor levels and enhances tissue repair, reduces musculoskeletal pain. Despite of uses for many years, the effect of dextrose solution on cellular and molecular base is not fully clear. Here, the roles of dextrose solutions was tried to find out in different concentrations on human fibroblasts in vitro. Gene expression alterations were analyzed in uses of dextrose solutions on growth and apoptotic factors. Methods:The effects of dextrose solution (1%, 5%, 10%-low doses, 15%, 20% and 25%-high doses) were evaluated in vitro by using human fibroblast culture. In each condition total RNA extraction and cDNA synthesis were performed. The gene expression levels of angiogenic and apoptotic factors were analyzed by using real-time polymerase chain reaction. The gene expression results of growth and apoptotic factors were correlated with control results. Results:Results;Dextrose solutions were affected the viability of fibroblast cells in culture flask in high concentrations. In high doses dextrose concentrations, up to 80% of fibroblasts were died because of toxic conditions. Viable fibroblast cell ratios were decreased proportionally due to the dextrose concentration. Low dextrose concentrations increased gene expressions in angiogenic (VEGFA, PDGFA, PDGFB, IGF1) and in apopitotic factors (CASP3 and CASP8) in fibroblasts. Conclusions: Conclusion; Dextrose solution in high concentrations, decreases viable cell ratios on adult fibroblast cell line. Dextrose solutions in proper concentrations increase the gene expressions of angiagenic and apopitotic factors on viable cells in adult fibroblast cell culture.

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Viability measurements of hybridoma cells in suspension cultures

Cited by 24 publications

References 20 publications

Effects of H2O2 on the growth, secretion, and metabolism of hybridoma cells in culture

Effects of H2O2 on the growth, secretion, and metabolism of hybridoma cells in culture

Dynamic Recording of Cell Death in the In Vitro Dorsal Vagal Nucleus of Rats in Response to Metabolic Arrest

Dextrose solution used for prolotherapy decreases cell viability and increases gene expressions of angiogenic and apopitotic factors

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