13 research outputs found
Network Glasses Under Pressure:Permanent Densification in Modifier-Free Al2O3−B2O3−P2O5−SiO2 Systems
Chemical Identity and Mechanism of Action and Formation of a Cell Growth Inhibitory Compound from Polycarbonate Flasks
This paper reports the chemical identity and mechanism of action and formation of a cell growth inhibitory compound leached from some single-use Erlenmeyer polycarbonate shaker flasks under routine cell culture conditions. Single-use cell culture vessels have been increasingly used for the production of biopharmaceuticals; however, they often suffer from issues associated with leachables that may interfere with cell growth and protein stability. Here, high-performance liquid-chromatography preparations and cell proliferation assays led to identification of a compound from the water extracts of some polycarbonate flasks, which exhibited subline- and seeding density-dependent growth inhibition of CHO cells in suspension culture. Mass spectroscopy, nuclear magnetic resonance spectroscopy, and chemical synthesis confirmed that this compound is 3,S-dinitro-bisphenol A. Cell cycle analysis suggests that 3,5-dinitro-bisphenol A arrests CHO-S cells at the G(1)/G(0) phase. Dynamic mass redistribution assays showed that 3,5-dinitro-bisphenol A is a weak GPR3S agonist. Analysis of the flask manufacturing process suggests that 3,5-dinitro-bisphenol A is formed via the combination of molding process with gamma-sterilization. This is the first report of a cell culture/assay interfering leachable compound that is formed through gamma-irradiation-mediated nitric oxide free radical reaction
Chemical Identity and Mechanism of Action and Formation of a Cell Growth Inhibitory Compound from Polycarbonate Flasks
This paper reports
the chemical identity and mechanism of action
and formation of a cell growth inhibitory compound leached from some
single-use Erlenmeyer polycarbonate shaker flasks under routine cell
culture conditions. Single-use cell culture vessels have been increasingly
used for the production of biopharmaceuticals; however, they often
suffer from issues associated with leachables that may interfere with
cell growth and protein stability. Here, high-performance liquid-chromatography
preparations and cell proliferation assays led to identification of
a compound from the water extracts of some polycarbonate flasks, which
exhibited subline- and seeding density-dependent growth inhibition
of CHO cells in suspension culture. Mass spectroscopy, nuclear magnetic
resonance spectroscopy, and chemical synthesis confirmed that this
compound is 3,5-dinitro-bisphenol A. Cell cycle analysis suggests
that 3,5-dinitro-bisphenol A arrests CHO-S cells at the G<sub>1</sub>/G<sub>o</sub> phase. Dynamic mass redistribution assays showed that
3,5-dinitro-bisphenol A is a weak GPR35 agonist. Analysis of the flask
manufacturing process suggests that 3,5-dinitro-bisphenol A is formed
via the combination of molding process with γ-sterilization.
This is the first report of a cell culture/assay interfering leachable
compound that is formed through γ-irradiation-mediated nitric
oxide free radical reaction