eoe sub articles

EoE Sub Articles

All procedures involving sample collection have been performed in accordance with the institute&#39;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 Mycobacterium tub...

agarose-based preparation of human precision-cut lung slices

Source: Neuhaus, V. et al., <a href="https://app.jove.com/v/57042">Assessment of the Cytotoxic and Immunomodulatory Effects of Substances in Human Precision-cut Lung Slices.</a> J. Vis. Exp. (2018)
In this video, we describe a procedure to inflate isolated human lungs with agarose and prepare human precision-cut lung slices. The prepared ex vivo lung slices can be used for further immunologic experiments.

Agarose-Based Preparation of Human Precision-Cut Lung Slices

All procedures involving sample collection have been performed in accordance with the institute&#39;s IRB guidelines.
1. General Preparation of Human PCLS ...

Begin with a resected human lung.
Insert a silicone tube into the main bronchus of the lung. Secure the tube, providing a pathway for introducing agarose into the tissue in the subsequent steps.&#160;
Clamp other bronchi and vessels to prevent agarose leakage during filling.
Introduce pre-warmed low-gelling agarose mixed with media into the lung until it is fully inflated.&#160;
Place the lung on ice for agarose polymerization, forming a solid gel matrix that provides stability to the tissue and maintains lung architecture during sectioning procedures.
After polymerization, cut the lung tissue into uniform slabs. Extract cylindrical tissue cores from the tissue slabs.
Using a tissue slicer filled with chilled buffer, slice the tissue core to obtain precision-cut lung slices.
Transfer the slices to a culture plate containing chilled media. Incubate to stabilize the tissue.
Finally, transfer the slices into a multi-well plate containing media. The prepared precision-cut lung slices are now ready for experiments.