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Live reporting for hypoxia: Hypoxia sensor–modified mesenchymal stem cells as in vitro reporters
Author(s) -
Schmitz Carola,
Pepelanova Iliyana,
Seliktar Dror,
Potekhina Ekaterina,
Belousov Vsevolod V.,
Scheper Thomas,
Lavrentieva Antonina
Publication year - 2020
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.27503
Subject(s) - mesenchymal stem cell , microbiology and biotechnology , hypoxia (environmental) , flow cytometry , stem cell , adipose tissue , tissue engineering , 3d cell culture , cell culture , chemistry , in vitro , cell , biophysics , biology , biomedical engineering , oxygen , immunology , biochemistry , medicine , genetics , organic chemistry
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% O 2 ). 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.