Assessment of long-term cultivated human precision-cut lung slices as an ex vivo system for evaluation of chronic cytotoxicity and functionality

Abstract

BackgroundInvestigation of basic chronic inflammatory mechanisms and development of new therapeutics targeting the respiratory tract requires appropriate testing systems, including those to monitor long- persistence. Human precision-cut lung slices (PCLS) have been demonstrated to mimic the human respiratory tract and have potential of an alternative, ex-vivo system to replace or augment in-vitro testing and animal models. So far, most research on PCLS has been conducted for short cultivation periods (≤72 h), while analyses of slowly metabolized therapeutics require long-term survival of PCLS in culture. In the present study, we evaluated viability, physiology and structural integrity of PCLS cultured for up to 15 days.MethodsPCLS were cultured for 15 days and various parameters were assessed at different time points.ResultsStructural integrity and viability of cultured PCLS remained constant for 15 days. Moreover, bronchoconstriction was inducible over the whole period of cultivation, though with decreased sensitivity (EC501d = 4 × 10−8 M vs. EC5015d = 4 × 10−6 M) and reduced maximum of initial airway area (1d = 0.5% vs. 15d = 18.7%). In contrast, even though still clearly inducible compared to medium control, LPS-induced TNF-α secretion decreased significantly from day 1 to day 15 of culture.ConclusionsOverall, though long-term cultivation of PCLS need further investigation for cytokine secretion, possibly on a cellular level, PCLS are feasible for bronchoconstriction studies and toxicity assays.Electronic supplementary materialThe online version of this article (doi:10.1186/s12995-017-0158-5) contains supplementary material, which is available to authorized users.

21st Century Tools for Nanotoxicology: Transcriptomic Biomarker Panel and Precision‐Cut Lung Slice Organ Mimic System for the Assessment of Nanomaterial‐Induced Lung Fibrosis

“…This study demonstrated that the thin 250 µm slices maintained structural integrity for 48 h; whereas, the thicker 500 µm slices were structurally intact in cultures up to 7 days postharvesting. This is different to what was shown previously by Neuhaus et al, [67] where increasing slice thickness decreased Figure 7. A) Schematic heat map depicting fold changes of 23 cytokine expression via 23-plex cytokine ELISA in supernatant from 250 µm PCLS exposed to 25, 50, and 100 µg mL −1 of Mitsui-7 compared to that of 250 µm PCLS exposed to media control on day 1 post-exposure.…”

A High-Throughput System for Cyclic Stretching of Precision-Cut Lung Slices During Acute Cigarette Smoke Extract Exposure

“…PCLSs also benefit by preserving the native extracellular environment (Sanderson, 2011) and retaining nearly all of the resident cell types in the lung. These technical advantages have thus promoted widespread adoption of the PCLS in models of exposure assessment (Langer et al, 2012;Lauenstein et al, 2014;Uhl et al, 2015;Hess et al, 2016;Watson et al, 2016;Neuhaus et al, 2017), pharmacologic therapy (Switalla et al, 2010;van Rijt et al, 2015;Donovan et al, 2016;Kistemaker et al, 2017), and disease modeling, including chronic obstructive pulmonary disease (COPD) (Chronic Obstructive Lung Disease [COLD], 2017).…”

Where We Stand: Lung Organotypic Living Systems That Emulate Human-Relevant Host–Environment/Pathogen Interactions

“…Besides, the PCLS cannot fully recapitulate all attributes seen in vivo; for instance, there are major challenges with preserving arteriole morphology and localization of lung airway smooth muscle cells (Sanderson, 2011). Precision cut lung slices can reproduce the initial interactions and inflammatory responses to industrial chemicals (Lauenstein et al, 2014) and pathogens [e.g., influenza viruses (Liu et al, 2015), rhinovirus (Beale et al, 2014), Yersinia pestis (Banerjee et al, 2019), and Coxiella burnetiid (Graham et al, 2016)] in the human respiratory system; however, the extent to which the immune response can replicate the in vivo situation is limited, as demonstrated by Neuhaus et al (2017), who observed that LPS-induced tumor necrosis factor-alpha (TNF-α) secretion decreased significantly over a period of 15 days, and it is not possible to recruit non-resident immune cells. Moreover, while murine-derived PCLS models have been adapted to study the effects of cigarette smoke (Donovan et al, 2016), the PCLS per se have not been utilized for inhalation exposure studies in vitro, principally because the route of administration represents a challenge (e.g., physiological ALI cannot be established in this model system), and often the entire slice is bathed in the compound or stimulant of interest (Liu et al, 2019).…”