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…

Divulgaciones

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

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Citar este artículo
Evaluating Cytotoxicity and Membrane Integrity in Lung Slices via Lactate Dehydrogenase Assay. J. Vis. Exp. (Pending Publication), e21849, doi: (2023).

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