This study outlines a method for establishing a mouse model to investigate lung cancer cell dissemination in the pleural cavity. The process involves injecting cancer cells into immunodeficient mice and observing the resulting tumor formation and pleural effusion.
To begin, prepare an immunodeficient mouse.
Using an assembly with a bent needle tip, inject a lung cancer cell suspension into the thoracic cavity.
The cancer cells reach the pleura containing a small amount of pleural fluid.
These cancer cells' receptors bind to specific ECM polysaccharides produced by the mesothelial cells lining the pleura, facilitating cell attachment to the pleural surface.
The cancer cells process these polysaccharides, causing increased cancer cell migration toward the surface. They also release proteolytic enzymes and autocrine growth factors.
Proteolytic enzymes degrade the ECM proteins, promoting cancer cell invasion.
The absence of a functional immune system and the presence of autocrine growth factors, allow the unrestricted proliferation of the cancer cells, forming tumor nodules.
Over time, tumor nodules grow and disseminate within the pleural cavity, disrupting the normal physiological fluid balance.
This results in fluid accumulation in the pleural cavity, a condition known as pleural effusion.
To set up a mouse dissemination model, use polystyrene foam to make a stopper, and disinfect it with 70% ethanol. Attach the needle to the stopper with the tip extending 5 millimeters out of the stopper. Bend the tip of a 30-gauge needle with clean forceps or against a hard object cleaned with 70% ethanol, and fill a syringe with 1 x 106 target tumor cells in 100 microliters of PBS.
Confirm a lack of response to pedal reflex in an anesthetized 19- to 21-gram 8- to 12-week-old female homozygote athymic nude mouse. Insert the needle into the chest through the intercostal space, moving the needle up and down to avoid contacting the ribs. When the tip has passed through the intercostal space, position the syringe so that it is pressed against the mouse and eject the entire volume of target cells. After the injection, roll the mouse 2 to 3 times to spread the cells throughout the thoracic cavity and return the mouse to its cage with monitoring until full recovery.
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