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Two of the most commonly used productivity-enhancing tools in CHO cell cultures during biopharmaceutical production are low temperature and sodium butyrate. While these two strategies change the balance in cell growth and productivity's reciprocal relationship, we do not fully know their mechanisms of action beyond a gross cell growth inhibition. Here, we used continuous culture to investigate the effect of low temperature and NaBu supplementation on CHO cell stability and gene expression profiles. According to a transcriptome analysis, the molecular mechanisms by which low temperature and NaBu arrested cell cycle in G1/G0 differed from each other by deregulation of different cell cycle checkpoints and regulators differed from each other. The results presented here provide new molecular evidence regarding cell cycle regulation during CHO cell bioprocessing and its implications for increased recombinant protein production.
Source link: https://doi.org/10.1371/journal.pone.0277620
Osteoclasts are the only singlenucleated cells in vivo responsible for bone resorption and are essential for regulating bone remodeling and bone mass maintenance. We investigated the effect of culture conditions on osteoclast differentiation, including cell density and receptor activator of nuclear factor kappa-B ligand concentrations with or without macrophage colony-stimulating factors. cytoplasmic 1 and cytoplasmic 1 were used to determine gene expression of osteoclast-specific gene marker cathepsin K, osteoclast transcription factors c-Fos and nuclear factor of activated T cells by using qPCR. RANKL therapy resulted in the generation of multinucleated osteoclasts and elevated CTSK, c-Fos, and NFATc1 gene expression, as well as increased RANKL-treated osteoclasts production and increased NFATc1 gene expression. M-CSF significantly reduced multinucleated osteoclasts formation, decreased CTSK gene expression, and had no effect on c-Fos and NFATc1 gene expression in comparison to RANKL treatment. Concerning bone resorption activity, RANKL's treatment increased bone resorption pits on bovine bone slices.
Source link: https://doi.org/10.1371/journal.pone.0277871
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