In Vivo Murine Liver Perfusion: A Method to Infuse the Rodent Liver with Solution via the Portal Vein
0 views • 3:18 min • April 30th, 2023
After anesthetizing and preparing the rodent, locate the portal vein and slide a thread under it. Tie a loose knot around the portal vein without pulling it taut. This will stabilize an IV catheter later during the protocol.
Holding the needle of an IV catheter bevel-up and parallel to the portal vein, insert it into the lumen - the space inside of the vein. Retract the needle and push the catheter through the vein all the way to the venous branched area within the liver.
After stabilizing the catheter, increase the peristaltic pump rate. Immediately, the liver should blanch as blood is replaced by perfusion liquid. Cut a major blood vessel to drain even more blood and perfusion liquid from the liver.
Squeeze the severed vessel a few times to inflate the liver and ensure it is completely devoid of blood. Switch the pump so that it perfuses buffer into the liver. Finally, repeat liver inflation briefly, taking care not to burst the liver from its connective tissue.
To place the portal vein catheter, first, use the back of a forceps to move the intestines to the right side of the mouse abdominal cavity, exposing the portal vein, and use curved forceps to slide a 20-centimeter piece of polyester sewing thread under the portal vein between the liver and the superior pancreaticoduodenal vein.
Use the forceps to carefully pull the thread to the other side of the animal so that it is centered under the portal vein and tie a loose overhead knot around the portal vein. Place the curved forceps under the vein and gently pull all tissue material toward the tail to straighten out the vein.
Next, place the bevel of a catheter needle facing up and parallel with the lower part of the portal vein near the forceps and gently puncture the vein with the needle. When the bevel is in the lumen of the vein, retract the spring-loaded needle and continue to push the polymer catheter through the vein until the bevel is near the venous branched area.
After securing the knot around the catheter, immediately set the peristaltic pump at 4 milliliters per minute and connect the male end of the tubing to the female end of the catheter, at which point you should start to see the liver blanch. Cut the aorta abdominalis for drainage. Use masking tape to immobilize the tubing onto the underpad.
Use the scissors to cut the abdominal skin vertically to allow drainage of blood and perfusion liquid. Then, squeeze the effluent blood vessel a few times with straight forceps to inflate the liver, ensuring that all of the blood has drained out, and perfuse the liver with PBS until the liver tissue blanches.
Overview
This article describes a detailed protocol for catheterizing the portal vein in rodents, focusing on the surgical techniques required for successful liver perfusion. The methodology includes anesthetization, catheter insertion, and perfusion procedures to ensure proper blood replacement in the liver.
Key Study Components
Area of Science
- Neuroscience
- Biology
- Surgical Techniques
Background
- Portal vein catheterization is crucial for liver studies.
- Proper techniques ensure accurate perfusion and tissue analysis.
- Understanding blood flow dynamics in the liver is essential for metabolic studies.
- Rodent models are commonly used in biomedical research.
Purpose of Study
- To establish a reliable method for portal vein catheterization.
- To facilitate liver perfusion for experimental purposes.
- To enhance the understanding of liver function and pathology.
Methods Used
- Anesthetization of the rodent.
- Identification and stabilization of the portal vein.
- Insertion of an IV catheter into the portal vein.
- Perfusion with buffer to replace blood in the liver.
Main Results
- The liver successfully blanched during perfusion.
- Blood drainage was effectively achieved through major blood vessels.
- Inflation of the liver was confirmed without tissue rupture.
- The protocol demonstrated reproducibility in catheter placement.
Conclusions
- The described method is effective for portal vein catheterization.
- It provides a foundation for further liver studies.
- Future research can build on this technique for metabolic investigations.
Frequently Asked Questions
Executive Industry Relevance
Portal vein catheterization enables precise delivery of compounds to the rodent liver, supporting target validation in metabolic disease models. This method provides a reproducible system for assessing hepatic uptake, clearance, and mechanism of action, contributing to predictive confidence in preclinical lead identification. By establishing a controlled perfusion environment, researchers can de-risk mechanistic hypotheses before advancing compounds into later-stage studies.
Strategic Applications in Biopharma R&D
Early Discovery & Target Validation
- Scientific Value: Enables interrogation of hepatic target engagement through controlled compound delivery via the portal vein.
- Operational Value: Provides a standardized surgical approach for consistent liver exposure across experimental cohorts.
Screening & Assay Development
- Scientific Value: Prepares the liver for perfusion-based assays that measure compound metabolism or biomarker release.
- Operational Value: Supports assay standardization by ensuring reproducible vascular access and perfusion conditions.
Translational & Preclinical Research
- Scientific Value: Maintains disease relevance by preserving native liver architecture and sinusoidal flow during perfusion.
- Operational Value: Facilitates continuity from discovery to preclinical validation through standardized tissue preparation.
Pipeline & Workflow Integration
This method integrates into the discovery continuum by enabling hepatic target validation after lead identification and prior to preclinical efficacy studies.
- Discovery Biology: Supports hypothesis testing by allowing direct infusion of compounds into the liver sinusoids.
- Screening: Enables assay readiness through reliable vascular cannulation and perfusion setup.
- Analytics: Facilitates quantitative measurement of hepatic extraction ratios or metabolite formation.
- Translational Research: Connects to preclinical work by providing perfused liver samples for downstream analysis.
- Enterprise Reuse: Establishes a reusable surgical platform for multiple hepatic perfusion experiments.
Operational & Enterprise Impact
- Scientific Value: Enhances target validation confidence by reducing variability in hepatic compound exposure.
- Operational Value: Improves reproducibility through standardized catheterization and perfusion protocols.
- Strategic Value: Informs go/no-go decisions by providing mechanistic data on liver-specific drug interactions.
- Portfolio Impact: Supports risk-adjusted prioritization of compounds based on hepatic clearance and metabolic stability.
Implementation Considerations
- Requires expertise in rodent vascular surgery and microsuturing techniques.
- Dependent on peristaltic pump and IV catheter infrastructure for controlled perfusion.
- Necessitates standardization across operators to ensure consistent venous access and flow rates.
- Must account for variations in rodent anatomy when adapting the technique across strains or ages.
- Limited by the need for terminal procedures, restricting longitudinal sampling in the same animal.
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Last updated: 22 August 2026