This research aims to enhance the rodents model for venous thrombosis, focusing on cancer-associated thrombosis. It introduces a refined surgical technique using vascular clips in a syngenic colon cancer xenograft mouse model to improve consistency and reproducibility. Our vast cancer associated thrombosis mechanistic understanding, and drug discovery.
Although the flow reduction model was introduced in the IVC stenotic models in 2012, no significant progress has been made in cancer associated thrombosis models in the last 10 years. This was the rationale for this endeavor. In the field of cancer associated thrombosis research, current technologies include advanced imaging techniques like scanning electron microscopic images, molecular biology tools such as immunofluorescence staining, precision surgical instrument, data analysis software and data interpretation, And animal welfare.
The current experimental challenges include variability in claw waste, a high mortality rate, and a prolonged learning curve in rodent models of venous thrombosis. Additionally, the venous flow direction and clot formation mechanisms differ from clinical cases. Our research protocol bridges gaps in rodent models of cancer associated thrombosis.
It introduces a novel surgical technique using vascular clips in xenograft models, yielding more consistent and reproducible thrombosis outcomes. This enhances our understanding of cancer related thrombosis and drug discovery in translational research. To begin, use a pair of fine scissors to make an incision on the abdominal skin of an anesthetized mouse, starting from the zyphoidal eminence to the bladder.
With a pair of atraumatic forceps, pinch the peritoneum halfway along the skin incision, ensuring an adequate gap between the incision and the overlying intestines. Next, open the peritoneum longitudinally along the avascular linea alba. Then use wet cotton tips to pull out the intestines carefully.
Then drop 200 to 300 microliters of normal saline over the intestines, and explore the route of the ureters inferior vena cava or IVC, aorta and renal veins. Cauterize the bilateral branches distal to the confluence of the left renal vein and infrarenal inferior vena cava. Check the ureters and vasculature for any oozing.
To prevent potential complications of spinal ischemia and claudication, leave the lumbar branches intact. Check for the side branches, including the bilateral uterine horn and hypogastric vessels. Next, use a closed tip atraumatic forceps to gently pass a six zero polypropylene suture through the avascular plane, two to three times.
Apply gentle pressure with a cotton tip tapinate to control any oozing. Finally, pass a five zero polypropylene suture through the plane with gentle coddled to cranial movements to widen it. Place the vascular clamps over the suture to guarantee adequate space.
Clot weights of the xenograft experimental group displayed a 1.5 fold increase relative to the control mice. Immunofluorescence assays showed that the IVC from the control group had intact CD 31 and fibrin partially occupying the vessel lumen. The CD 31 expression was not intact in the experimental group with the wall showing fibrin staining.
The control group had significantly lower fibrin expression compared to the experimental group. Ultrasonic graphic analysis of the mice showed the ligated infrarenal vena cava of a control group mouse had a partial clot. The experimental group, however, had a large clot, nearly occluding the infrarenal vena cava.
