Live reporting for hypoxia: Hypoxia sensor–modified mesenchymal stem cells as in vitro reporters

Carola Schmitz, Iliyana Pepelanova, Dror Seliktar, Ekaterina Potekhina, Vsevolod V. Belousov, Thomas Scheper, Antonina Lavrentieva

Research output: Contribution to journalArticlepeer-review


Natural oxygen gradients occur in tissues of biological organisms and also in the context of three-dimensional (3D) in vitro cultivation. Oxygen diffusion limitation and metabolic oxygen consumption by embedded cells produce areas of hypoxia in the tissue/matrix. However, reliable systems to detect oxygen gradients and cellular response to hypoxia in 3D cell culture systems are still missing. In this study, we developed a system for visualization of oxygen gradients in 3D using human adipose tissue–derived mesenchymal stem cells (hAD-MSCs) modified to stably express a fluorescent genetically engineered hypoxia sensor HRE-dUnaG. Modified cells retained their stem cell characteristics in terms of proliferation and differentiation capacity. The hypoxia-reporter cells were evaluated by fluorescence microscopy and flow cytometry under variable oxygen levels (2.5%, 5%, and 7.5% O2). We demonstrated that reporter hAD-MSCs output is sensitive to different oxygen levels and displays fast decay kinetics after reoxygenation. Additionally, the reporter cells were encapsulated in bulk hydrogels with a variable cell number, to investigate the sensor response in model 3D cell culture applications. The use of hypoxia-reporting cells based on MSCs represents a valuable tool for approaching the genuine in vivo cellular microenvironment and will allow a better understanding of the regenerative potential of AD-MSCs.

Original languageEnglish
Pages (from-to)3265-3276
Number of pages12
JournalBiotechnology and Bioengineering
Issue number11
StatePublished - 1 Nov 2020


  • 3D cell culture
  • AD-MSCs
  • hydrogels
  • hypoxia sensor
  • reporter cells

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology


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