Assessment of the Cytotoxic and Immunomodulatory Effects of Substances in Human Precision-cut Lung Slices

Abstract

Respiratory diseases in their broad diversity need appropriate model systems to understand the underlying mechanisms and enable development of new therapeutics. Additionally, registration of new substances requires appropriate risk assessment with adequate testing systems to avoid the risk of individuals being harmed, for example, in the working environment. Such risk assessments are usually conducted in animal studies. In view of the 3Rs principle and public skepticism against animal experiments, human alternative methods, such as precision-cut lung slices (PCLS), have been evolving. The present paper describes the ex vivo technique of human PCLS to study the immunomodulatory potential of low-molecular-weight substances, such as ammonium hexachloroplatinate (HClPt). Measured endpoints include viability and local respiratory inflammation, marked by altered secretion of cytokines and chemokines. Pro-inflammatory cytokines, tumor necrosis factor alpha (TNF-α), and interleukin 1 alpha (IL-1α) were significantly increased in human PCLS after exposure to a sub-toxic concentration of HClPt. Even though the technique of PCLS has been substantially optimized over the past decades, its applicability for the testing of immunomodulation is still in development. Therefore, the results presented here are preliminary, even though they show the potential of human PCLS as a valuable tool in respiratory research.

Time‐Resolved Quantification of Nanoparticle Uptake, Distribution, and Impact in Precision‐Cut Liver Slices

“…Several studies that use tissue slices with nanoparticles have been published 11–14. In most cases, slices have been used to determine nanoparticle impact on the tissue, but also to test nanoparticle formulations for drug delivery, with many examples focused on the lungs,11,13,15–17 but also intestine,18 liver,1,2 and tumor tissue 19. Most of these works used tissue from laboratory animals, with the exception of one study using human tissue from the lungs17 and one from intestine 18.…”

Intraclonal competitive fitness of longitudinal cystic fibrosis Pseudomonas aeruginosa airway isolates in liquid cultures

“…Hence, a sputum medium would have been an inappropriate choice for our competition experiments. However, to come closer to the habitat of the CF lungs, we consider to perform the next round of competitive fitness experiments in the ex vivo lung model of precision-cut lung slices (Neuhaus et al, 2018).…”

An antimicrobial molecule mitigates signs of sepsis in vivo and eradicates infections from lung tissue

“…35,36 Briefly, the trachea was cannulated and the lungs were inflated with 37°C warm, 1.5% low-gelling agarose medium solution (Sigma-Aldrich, Munich, Germany) and cooled on ice. Animals were housed under conventional housing conditions (22 ± 2°C, relative humidity 55 ± 15%, 12 hours dark/ light cycle).…”

“…35,36 Briefly, PCLSs were incubated in 4 µM solution of Calcein-AM in DMEM-F12 for 45 minutes at 37°C and 150 rpm in the dark. 35,36 Briefly, PCLSs were incubated in 4 µM solution of Calcein-AM in DMEM-F12 for 45 minutes at 37°C and 150 rpm in the dark.…”

“…PCLS were prepared as previously described. 35,36 Briefly, the trachea was cannulated and the lungs were inflated with 37°C warm, 1.5% low-gelling agarose medium solution (Sigma-Aldrich, Munich, Germany) and cooled on ice. After polymerization of the agarose, 8 mm wide tissue cores were prepared and cut into 200-to 300-µm-thick slices using a Krumdieck microtome (Alabama Research and Development, Munford, AL, USA) and collected in 4°C cold EBSS (Sigma-Aldrich, Munich, Germany).…”