An Assay to Measure Chemically-Induced Cytotoxicity in Human Precision-Cut Lung Slices

Published: November 30, 2023

Abstract

Source: Neuhaus, V. et al., Assessment of the Cytotoxic and Immunomodulatory Effects of Substances in Human Precision-cut Lung Slices. J. Vis. Exp. (2018)

This video illustrates a technique for evaluating cytotoxicity in human precision-cut lung slices through the WST-1 assay. Human precision-cut lung slices are subjected to incremental concentrations of a cytotoxic chemical to quantify their cytotoxicity. A decrease in color intensity serves as confirmation of the chemical's detrimental impact on lung cells.

Protocol

All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines. 1. General Preparation of Human precision-cut lung slices (PCLS) and Subsequent Exposure to Chemicals NOTE: Two persons are required to fill a lung. An up-to-date vaccination record for hepatitis A and B is recommended. Patients are routinely screened for hepatitis C virus (HCV) and human immunodeficiency virus (HIV) p…

Divulgaciones

The authors have nothing to disclose.

Materials

Silicon hose 3.0 x 5.0 mm A. Hartenstein (Leipzig, Germany) SS04
Syringe Faust Lab Science (Klettgau, Germany) 9.410 050
Coring tools custom-made custom-made
Trimming Blade Handle (FEATHER) pfm medical ag  (Cologne, Germany) 205530001
Trimming Blades (FEATHER) pfm medical ag  (Cologne, Germany) 205500000
Microtome: Tissue Slicer Alabama R&D (Bad Homburg, Germany) 303400-ADPT Krumdieck Tissue Slicer (MD6000)
Microtome blade Wilkinson Sword (Solingen, Germany) ENR-4027800011506
Cell strainer filter (100 µm Nylon) Becton Dickinson (Heidelberg, Germany) BD352360
Inoculation loop Copan Diagnostics (Murrieta, USA)  CD176S01
TPP Tissue culture plates 24wells Sigma (München, Germany) Z707791-126EA
Nunc MaxiSorp flat-bottom Fisher Scientific GmbH (Hannover, Germany) 44-2404-21
Nunc MicroWell 96-Well  Thermo Scientific (Schwerte, Germany) 260836
Multiwell-Reader Tecan Group Ltd. (Männedorf, Switzerland) Tecan infinite F200Pro Plate Reader
Plate shaker Edmund Buehler GmbH (Hechingen, Germany) KM-2 Akku
Assays
Cell Proliferation Reagent WST-1 Roche (Basel, Switzerland) 11644807001
Reagents
Agarose, low gelling temperature  Sigma (München, Germany) A9414-100G
Balanced Salt Mixtures and Solutions, Cell Culture, Classic Media and Salts, Earle's Balanced Salts (EBSS) Sigma (München, Germany) E2888-500ML
Penicillin and streptomycin Lonza (Verviers, Belgium) 17-602E
Dulbecco´s Modified Eagle´s Medium Nutrient Mixture F-12 Ham (DMEM F-12) Gibco (Darmstadt, Germany) 11039-047 Culture medium
Dulbecco´s Phosphate with Ca and Mg (DPBS) Lonza (Verviers, Belgium) BE17-513F   Buffer solution
Detergent Sigma (Saint Louis, USA) X100-100ML Triton X-100

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Citar este artículo
An Assay to Measure Chemically-Induced Cytotoxicity in Human Precision-Cut Lung Slices. J. Vis. Exp. (Pending Publication), e21848, doi: (2023).

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