The effects of oxygen concentration on cell death, anti-oxidant transcription, acute inflammation, and cell proliferation in precision-cut lung slices

Mitchel J R Ruigrok,Henderik W Frijlink,Peter Olinga,Jasmine Tomar,Wouter L J Hinrichs,Barbro N Melgert

doi:10.1038/s41598-019-52813-2

Mitchel J R Ruigrok, Henderik W Frijlink + Show 4 more

Open Access

https://doi.org/10.1038/s41598-019-52813-2

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Journal: Scientific Reports	Publication Date: Nov 7, 2019
Citations: 12	License type: open-access

Affiliation: University of Groningen

Abstract

Although animal models are often used in drug research, alternative experimental models are becoming more popular as they reduce animal use and suffering. Of particular interest are precision-cut lung slices, which refer to explants – with a reproducible thickness and diameter – that can be cultured ex vivo. Because lung slices (partially) reflect functional and structural features of whole tissue, they are often applied in the field of immunology, pharmacology, toxicology, and virology. Nevertheless, previous research failed to adequately address concerns with respect to the viability of lung slices. For instance, the effect of oxygen concentration on lung slice viability has never been thoroughly investigated. Therefore, the main goal of this study was to investigate the effect of oxygen concentration (20 vs. 80% O2) on the degree of cell death, anti-oxidant transcription, acute inflammation, and cell proliferation in lung slices. According to the results, slices incubated at 20% O2 displayed less cell death, anti-oxidant transcription, and acute inflammation, as well as more cell proliferation, demonstrating that these slices were considerably more viable than slices cultured at 80% O2. These findings expand our knowledge on lung slices and their use as an alternative experimental model in drug research.

Highlights

Animal models are often used in drug research, alternative experimental models are becoming more popular as they reduce animal use and suffering
To check whether caspase-dependent apoptosis was triggered in slices, we examined Casp[3] mRNA expression and accumulation of its functional protein cl-CASP3 (Fig. 4)
Lung slices cultured at 20% O2 displayed less cell death, anti-oxidant transcription, and acute inflammation, as well as more cell proliferation, demonstrating these slices were significantly more viable than slices incubated at 80% O2

Summary

Introduction

Animal models are often used in drug research, alternative experimental models are becoming more popular as they reduce animal use and suffering. Slices incubated at 20% O2 displayed less cell death, antioxidant transcription, and acute inflammation, as well as more cell proliferation, demonstrating that these slices were considerably more viable than slices cultured at 80% O2 These findings expand our knowledge on lung slices and their use as an alternative experimental model in drug research. Researchers could use precision-cut lung slices, which are explants – with a reproducible thickness and diameter – that can be cultured ex vivo[1] Advantages of this model include its ability to (partially) reflect the functional and structural heterogeneity of whole tissue and to decrease animal use as more experimental conditions can be tested per animal. Cell proliferation in slices was assessed by measuring mRNA expression of proliferation-related genes and by staining lung slices for Ki67 – a marker of proliferation[23]

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