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