Lung-tumor Tissue Explant Culture: A Procedure to Generate and Culture Precision-cut Lung Tumor Slices

- Place freshly harvested tumor-bearing lungs on a tissue culture plate. Separate the lung lobes. Transfer a tumor-bearing lobe onto a filter paper to avoid slipping. Cut a part of the lobe surrounding the tumor to generate a flat surface. Immerse the flat surface in cyanoacrylate glue to facilitate adhesion.

Next, mount on a vibratome specimen holder, securing the tumor in an upright position. Transfer the specimen holder onto a buffer tray and fill the tray with a suitable buffer. Place the tray in an ice bath to prevent tissue damage from heat generated during slicing. Use a vibratome to obtain thin tumor tissue slices.

Simultaneously, assemble a customized titanium grid in a culture plate containing a suitable growth medium to support tissue slice culture. Angle the plate to allow a portion of the solution to cover the grid. Place the tissue slices in the medium. Transfer the plate into a rotating incubation unit. During rotating incubation, the tissue slices alternate between air and liquid phases, enabling proper exposure to nutrients and oxygen. In this protocol, we will demonstrate the preparation and subsequent culturing of lung tumor explants.

- Transfer the tumor-bearing