Reliable monitoring of mammalian cells in bioreactors is essential to biopharmaceutical production. Trypan blue exclusion is a method of determining cell density and viability that has been used for over one hundred years to monitor cells in culture and is the current standard method in biomanufacturing. This method has many disadvantages however and there is a growing demand for more detailed and in-line measurements of cell growth in bioreactors.
View Article and Find Full Text PDFThe manufacturing of recombinant protein is traditionally undertaken in mammalian cell culture. Today, speed, cost and safety are the primary considerations for process improvements in both upstream and downstream manufacturing. Leaders in the biopharmaceutical industry are striving for continuous improvements to increase throughput, lower costs and produce safer more efficacious drugs.
View Article and Find Full Text PDFA sensitive electrochemical immunoassay (e-ELISA) has been developed for the detection of the gastrointestinal parasitic nematode Ostertagia ostertagi (brown stomach worm) in infected and control serum samples. An antigen-indirect immunoassay format was employed to detect the presence of O. ostertagi antibodies, coupled with an anti-species monoclonal horseradish peroxidase (HRP) conjugate.
View Article and Find Full Text PDFThe rise of three-dimensional cell culture systems that provide in vivo-like environments for pharmaco-toxicological models has prompted the need for simple and robust viability assays suitable for complex cell architectural structures. This study addresses that challenge with the development of an in vitro enzyme based electrochemical sensor for viability/cytotoxicity assessment of two-dimensional (2D) monolayer and three-dimensional (3D) spheroid culture formats. The biosensor measures the cell viability/toxicity via electrochemical monitoring of the enzymatic activity of nonspecific esterases of viable cells, through the hydrolysis of 1-naphthyl acetate to 1-naphthol.
View Article and Find Full Text PDFIn this paper we report the effects of the complex III inhibitor, strobilurin fungicide kresoxim-methyl, on the cellular homeostasis of a mammalian cancerous neural cell line. We examined whether exposure to subcytotoxic concentrations of kresoxim-methyl induce cellular and biochemical mechanisms of toxicity on the murine neuroblastoma N2a cells. Results revealed elevation of mitochondrial superoxide generation, decrease in mitochondrial transmembrane potential, losses on GPx enzyme activity, along with increased nitrite release.
View Article and Find Full Text PDFBiochim Biophys Acta Gen Subj
September 2017
Background: The rise of organic electronics represents one of the most prominent technological developments of the last two decades, with its interface with biological systems highlighting new directions of research. The "soft" nature of conducting polymers renders them unique platforms for cell-based microdevices, allowing their implementation in drug discovery, pharmaceutical effect analysis, environmental pollutant testing etc.
Methods: Cellular adhesion, proliferation and viability experiments were carried out to verify the biocompatibility of a PEDOT conductive polymer surface.
In this paper we elucidate the effects of the cytochrome bc1 inhibitor, strobilurin fungicide kresoxim-methyl, on the redox balance of a mammalian renal cell line. To explore whether mammalian exposure to sub-nephrotoxic concentrations of kresoxim-methyl induces cellular and biochemical mechanisms of toxicity, its effects on cellular viability and, in particular, several parameters related to oxidative stress, mitochondrial respiratory function and apoptosis were examined in fibroblast-like renal Vero cells. Elevation of mitochondrial superoxide generation, together with a concomitant decrease in mitochondrial transmembrane potential was indicative of mitochondrial dysfunction.
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