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Stabilization of enzyme-immobilized hydrogels for extended hypoxic cell culture

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Abstract

In 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.

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Funding

This work was supported in part by a National Science Foundation Faculty Early Career Development (CAREER) Award (#1452390) and Walther Cancer Foundation Oncology Physical Sciences & Engineering Research Embedding Program.

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Authors and Affiliations

  1. Department of Biomedical Engineering, Indiana University-Purdue University Indianapolis, 723 W. Michigan St. SL220K, Indianapolis, IN, 46202, USA

    Britney N. Hudson, Camron S. Dawes, Nathan DImmitt & Chien-Chi Lin

  2. Weldon School of Biomedical Engineering, Purdue University, West Lafayette, IN, 47907, USA

    Hung-Yi Liu & Chien-Chi Lin

  3. Department of Medicine, Division of Hematology/Oncology, Indiana University School of Medicine, Indianapolis, IN, 46202, USA

    Fangli Chen & Heiko Konig

  4. Indiana University Melvin & Bren Simon Cancer Center, Indianapolis, IN, 46202, USA

    Heiko Konig & Chien-Chi Lin

Authors
  1. Britney N. Hudson
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  2. Camron S. Dawes
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  5. Fangli Chen
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  7. Chien-Chi Lin
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Correspondence to Chien-Chi Lin.

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Hudson, B.N., Dawes, C.S., Liu, HY. et al. Stabilization of enzyme-immobilized hydrogels for extended hypoxic cell culture. emergent mater. 2, 263–272 (2019). https://doi.org/10.1007/s42247-019-00038-4

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  • DOI: https://doi.org/10.1007/s42247-019-00038-4

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