Hudson, Britney N.Dawes, Camron S.Liu, Hung-YiDImmitt, NathanChen, FangliKonig, HeikoLin, Chien-Chi2024-02-012024-02-012019Hudson BN, Dawes CS, Liu HY, et al. Stabilization of enzyme-immobilized hydrogels for extended hypoxic cell culture. Emergent Mater. 2019;2(2):263-272. doi:10.1007/s42247-019-00038-4https://hdl.handle.net/1805/38252In this work, glucose oxidase (GOx)-immobilized hydrogels are developed and optimized as an easy and convenient means for creating solution hypoxia in a regular incubator. Specifically, acrylated GOx co-polymerizes with poly(ethylene glycol) diacrylate (PEGDA) to form PEGDA-GOx hydrogels. Results show that freeze-drying and reaction by-products, hydrogen peroxide, negatively affect oxygen-consuming activity of network-immobilized GOx. However, the negative effects of freeze-drying can be mitigated by addition of trehalose/raffinose in the hydrogel precursor solution, whereas the inhibition of GOx caused by hydrogen peroxide can be prevented via addition of glutathione (GSH) in the buffer/media. The ability to preserve enzyme activity following freeze-drying and during long-term incubation permits facile application of this material to induce long-term solution/media hypoxia in cell culture plasticware placed in a regular CO2 incubator.en-USPublisher PolicyHypoxiaCancerEnzyme immobilizationGlucose oxidaseHydrogelArticle