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Updated: May 9, 2026
Ex Vivo Hepatic Perfusion Through the Portal Vein in Mouse
Published on: March 9, 2022
Murine portal vein catheterization to analyze liver-directed therapies
Joseph D Valentino1, Piotr G Rychahou, W Conan Mustain
1Department of Surgery, University of Kentucky, Lexington, Kentucky; Markey Cancer Center, University of Kentucky, Lexington, Kentucky.
Background:
Small interfering RNA (siRNA) provides a highly selective method to target mutated pathways; however, its use is complicated by specific delivery to tumor cells. The aims of the present study were to develop a novel murine model of portal vein catheterization for the chronic delivery of therapeutic agents to liver metastases, determine the benefits of local delivery of siRNA to liver metastases, and determine the utility of epithelial cell adhesion molecule (EpCAM) as a selective target for siRNA delivery to colorectal cancer (CRC) metastases.
Materials And Methods:
First, portal vein catheterization was performed through a midline laparotomy in 2 mo-old Balb/C mice. Second, the portal venous flow distribution and catheter patency were evaluated using fluorescent-labeled microspheres. Metastatic studies were performed by splenic injection of CT26 murine colon cancer cells. Uptake of DY-547-labeled siRNA was assessed by IVIS imaging, with delivery to the metastases confirmed using fluorescent microscopy. Finally, EpCAM expression was evaluated using immunohistochemical staining of human tissue microarrays.
Results:
Successful portal vein catheterization was confirmed by saline injection and ultrasound. Fluorescent imaging of microspheres confirmed excellent distribution and catheter patency. Portal venous injection of DY547-labeled siRNA demonstrated a high level of fluorescence throughout the liver, with siRNA also identified within the liver metastases. Also, all primary CRCs and liver metastases stained strongly for EpCAM, with no expression in normal hepatocytes.
Conclusions:
Liver-directed therapy can provide the selective delivery of siRNA to CRC metastases. EpCAM expression in CRC, but not normal liver, could further selectively target hepatic metastases of epithelial origin.
Insights
Local delivery of small interfering RNA (siRNA) via portal vein catheterization effectively targets liver metastases. Epithelial cell adhesion molecule (EpCAM) serves as a selective target for colorectal cancer (CRC) liver metastases.
Area of Science:
- Oncology
- Molecular Biology
- Surgical Innovation
Background:
- Small interfering RNA (siRNA) offers targeted therapy but faces delivery challenges to tumor cells.
- Developing effective delivery methods for siRNA to liver metastases is crucial for cancer treatment.
- Identifying specific molecular targets on cancer cells enhances therapeutic selectivity.
Purpose of the Study:
- To establish a murine model for chronic delivery of therapeutics to liver metastases via portal vein catheterization.
- To evaluate the advantages of localized siRNA delivery to liver metastases.
- To assess the potential of epithelial cell adhesion molecule (EpCAM) as a targeted delivery marker for colorectal cancer (CRC) liver metastases.
Main Methods:
- Portal vein catheterization was performed in mice.
- Distribution and patency of the catheter were assessed using fluorescent microspheres.
- Colorectal cancer cells were injected to create liver metastases models.
- siRNA uptake and delivery to metastases were visualized using fluorescent imaging and microscopy.
- EpCAM expression was analyzed in human tissue microarrays.
Main Results:
- Portal vein catheterization was successfully established with confirmed patency and excellent flow distribution.
- siRNA demonstrated high liver distribution and uptake within liver metastases.
- All primary CRCs and liver metastases showed strong EpCAM expression, while normal hepatocytes did not.
Conclusions:
- Liver-directed therapy via portal vein catheterization enables selective siRNA delivery to colorectal cancer liver metastases.
- EpCAM expression in colorectal cancer, but not in normal liver cells, provides a specific target for hepatic metastases of epithelial origin.
