Evaluating Cytotoxicity and Membrane Integrity in Lung Slices via Lactate Dehydrogenase Assay
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 demonstrates the lactate dehydrogenase, LDH assay to measure the cytotoxic effect of a test substance and its effect on the membrane integrity in human precision-cut lung slices. The assay provides a quantitative measure of cell death or damage in the sample.
Protocol
All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines 1. General Preparation of Human 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 HCV and HIV prior to lung transplantation. If an active infection with M…
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The authors have nothing to disclose.
Materials
| Nunc MicroWell 96-Well | Thermo Scientific (Schwerte, Germany) | 260836 | |
| Image rendering software | Bitplane AG (Zürich, Switzerland) | IMARIS 7.6 | |
| LSM Image Browser | Zeiss (Jena, Germany) | ||
| Multiwell-Reader | Tecan Group Ltd. (Männedorf, Switzerland) | Tecan infinite F200Pro | Plate Reader |
| Cytotoxicity Detection Kit (LDH) | Roche (Basel, Switzerland) | 11644793001 | |
| Cell Proliferation Reagent WST-1 | Roche (Basel, Switzerland) | 11644807001 | |
| Washing buffer | Merck (Darmstadt Germany) | 524653 | 0.05% Tween 20 in PBS |
| Detergent | Sigma (Saint Louis, USA) | X100-100ML | Triton X-100 |
