Details
| Original language | English |
|---|---|
| Pages (from-to) | 3265-3276 |
| Number of pages | 12 |
| Journal | Biotechnology and bioengineering |
| Volume | 117 |
| Issue number | 11 |
| Early online date | 15 Jul 2020 |
| Publication status | Published - 15 Oct 2020 |
Abstract
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.
Keywords
- 3D cell culture, AD-MSCs, hydrogels, hypoxia sensor, reporter cells
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biotechnology
- Chemical Engineering(all)
- Bioengineering
- Immunology and Microbiology(all)
- Applied Microbiology and Biotechnology
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In: Biotechnology and bioengineering, Vol. 117, No. 11, 15.10.2020, p. 3265-3276.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Live reporting for hypoxia: Hypoxia sensor–modified mesenchymal stem cells as in vitro reporters
AU - Schmitz, Carola
AU - Pepelanova, Iliyana
AU - Seliktar, Dror
AU - Potekhina, Ekaterina
AU - Belousov, Vsevolod V.
AU - Scheper, Thomas
AU - Lavrentieva, Antonina
N1 - Funding information: This study was supported by the German Research Foundation (DFG Project 398007461 488 “3D Dual?Gradient Systems for Functional Cell Screening”) and Grant # 075?15?2019?1789 from the Ministry of Science and Higher Education of the Russian Federation. The publication of this article was funded by the Open Access Fund of Leibniz Universität Hannover. The authors also want to acknowledge support by the SMART BIOTECS initiative, financially supported by the Ministry of Science and Culture (MWK) of Lower Saxony, Germany. Open access funding enabled and organized by Projekt DEAL. This study was supported by the German Research Foundation (DFG Project 398007461 488 ?3D Dual-Gradient Systems for Functional Cell Screening?) and Grant # 075-15-2019-1789 from the Ministry of Science and Higher Education of the Russian Federation. The publication of this article was funded by the Open Access Fund of Leibniz Universit?t Hannover. The authors also want to acknowledge support by the SMART BIOTECS initiative, financially supported by the Ministry of Science and Culture (MWK) of Lower Saxony, Germany. Open access funding enabled and organized by Projekt DEAL.
PY - 2020/10/15
Y1 - 2020/10/15
N2 - 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.
AB - 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.
KW - 3D cell culture
KW - AD-MSCs
KW - hydrogels
KW - hypoxia sensor
KW - reporter cells
UR - http://www.scopus.com/inward/record.url?scp=85088565064&partnerID=8YFLogxK
U2 - 10.1002/bit.27503
DO - 10.1002/bit.27503
M3 - Article
C2 - 32667700
AN - SCOPUS:85088565064
VL - 117
SP - 3265
EP - 3276
JO - Biotechnology and bioengineering
JF - Biotechnology and bioengineering
SN - 0006-3592
IS - 11
ER -