A Retrograde Implantation Approach for Peritoneal Dialysis Catheter Placement in Mice
July 20th, 2022
This article describes modifications of a procedure to implant a peritoneal dialysis catheter in a murine model to avoid major technical issues observed with the conventional techniques.
The previous protocols for mouse models of peritoneal dialysis suffer from limitations precluding their long-term use. These limitations, such as catheter coiling and dislodgement, are addressed in this study. This technique prevents PD catheter malfunction for mechanical reasons, allowing their use for a longer time to study pathologies, such as peritoneal fibrosis.
This technique will provide a better mouse model to probe molecular pathogenesis and novel therapeutics of peritoneal fibrosis. A key limitation of PD in humans. PD is the mainstay of renal replacement therapy in pediatric patients with ESKD.
This optimized model will provide a better test bed to investigate all the issues with PD in humans. Generation of an animal model has a learning curve. This step-by-step video will assist an individual in implementing this technique to generate a durable model for peritoneal dialysis successfully.
To begin place the fully anesthetized eight to 12 weeks old C57 black/6J mouse in a left lateral position. After shaving the mouse as specified in the manuscript, disinfect the surgical area by exposing the right flank of the mouse to the heating blanket. Then place the sterile surgical drape over the mouse.
Assign the access port pocket one centimeter above the mouse tail. Hold the installation segment with the non-dominant index and thumb finger over the assigned area near the tail. Place the catheter above the skin, and estimate the place for the catheter's tube insertion within the abdominal cavity.
Then mark the assigned place for tube insertion, respecting the minimal bending of the tube near the anterior midline. Punch a side hole over the frame of the reservoir section with the mouse ear tagger. Make a horizontal one centimeter wide skin incision one centimeter above the tail.
Then bluntly dissect the subcutaneous plane from the underlying muscular layer to make a pouch for the catheter placement to ensure the installation port freely resides in the ideal port pocket. Past the 3-0 suture from the customized side hole. Fix the access port to the muscular bed by tightening the passed suture, keeping the tubing course cephalad.
Make a one centimeter incision over the formally marked area near the midline. Place a loose purse string suture with 4-0 round absorbable suture around the incised abdominal wall muscle. Make a one centimeter incision over the muscular layer close to the right midline and pass the proximal felt of the catheter inside the incision.
Pass the catheter tube through the prepared tract. Tighten the prepared purse string suture around the tube, while keeping the second felt outside the purse string over the muscular layer, and close the skin with 3-0 absorbable sutures. Ensure that the placed catheter is functional.
Check the function with a one milliliter syringe attached to the specific Huber needle for the port. Inject normal saline into the installation port. After confirming a smooth flow with a zero tolerance for resistance, close the skin incisions around the port reservoir with 3-0 absorbable sutures.
Enter the port with the Huber needle and inject 100 microliters of normal saline into the port to confirm the patent course. Then inject 200 microliters of lipopolysaccharide, followed by 100 microliters of saline for tube irrigation and ensure that there is no resistance. After seven days of LPS injections and two weeks of catheter implantation, plan for the peritoneal biopsy.
After confirming successful anesthesia, make a midline skin incision from bladder to sub xiphoid. Then perfuse the subfascial plane with cold PBS. Make sure the plane is completely dissected without disturbing the integrity of the peritoneum.
Analysis of the hematoxylin and eosin stained sections showed a substantial increase in the extracellular matrix, or ECM, in the subperitoneal space. The average ECM in the sub peritoneal space of the control mice was half of that in the LPS exposed mice. The Masson's Trichrome staining detects fibrosis, measured as intensity density normalized to the surface area.
Altered vascularity and widening of the subperitoneal space were analyzed using endothelial cell marker CD31, and measured as integrated density in randomly selected high-power field images. Compared to the control, LPS-induced mice showed a threefold increase in subperitoneal fibrosis, approximately eight to ninefold increase in the vascularity, and a twofold increase in the subperitoneal space, marked as SP, were observed. Place the catheter above the skin and mark the point for the catheter insertion, while avoiding the bending of the tube.
The insertion should be near the interior midline. Providing the more durable murine PD catheter placement method will allow the researchers to design and execute longer term experiments to probe and target peritoneal membrane failure.
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Overview
This article presents a modified procedure for implanting a peritoneal dialysis catheter in a murine model, addressing significant technical issues associated with conventional methods. The new technique enhances the longevity and functionality of the catheter, facilitating long-term studies of related pathologies.
Key Study Components
Area of Science
- Neuroscience
- Biology
- Medical Research
Background
- Previous protocols for peritoneal dialysis in mouse models have limitations.
- Common issues include catheter coiling and dislodgement.
- These limitations hinder long-term studies of peritoneal dialysis.
- Peritoneal fibrosis is a key pathology of interest in this research.
Purpose of Study
- To improve the implantation technique for peritoneal dialysis catheters.
- To enable long-term use of catheters in murine models.
- To facilitate research into the molecular pathogenesis of peritoneal fibrosis.
Methods Used
- Modification of catheter implantation procedures.
- Assessment of catheter functionality over extended periods.
- Evaluation of the impact on peritoneal dialysis efficacy.
- Analysis of related pathologies in murine models.
Main Results
- The modified technique significantly reduces catheter malfunction.
- Long-term studies are now feasible with the new implantation method.
- Enhanced understanding of peritoneal fibrosis is achievable.
- The study provides a better model for testing novel therapeutics.
Conclusions
- The new implantation technique addresses critical limitations of previous methods.
- This advancement allows for more reliable studies of peritoneal dialysis.
- It opens avenues for exploring therapeutic options for peritoneal fibrosis.
Frequently Asked Questions
Executive Industry Relevance
Reliable murine models for peritoneal dialysis are essential for translational research targeting peritoneal membrane failure, a major complication in end-stage kidney disease. This optimized retrograde catheter implantation approach addresses mechanical limitations, enabling longer-term studies of peritoneal inflammation and fibrosis. Enhanced model durability supports predictive confidence in preclinical evaluation of novel therapeutics and mechanistic hypotheses.
Strategic Applications in Biopharma R&D
Early Discovery & Target Validation
- Enables robust interrogation of peritoneal fibrosis mechanisms relevant to human disease.
- Supports functional validation of molecular targets implicated in peritoneal membrane failure.
- Facilitates biological de-risking by providing a stable in vivo system for hypothesis testing.
- Improves predictive confidence for downstream therapeutic development decisions.
Screening & Assay Development
- Provides a validated murine platform for quantitative assessment of peritoneal inflammation and fibrosis.
- Enables reproducible induction and measurement of disease-relevant phenotypes using standardized catheter placement.
- Supports assay development for evaluating anti-fibrotic or anti-inflammatory interventions in vivo.
- Improves screening readiness by minimizing technical failures and animal attrition.
Translational & Preclinical Research
- Aligns preclinical model outputs with clinical endpoints such as peritoneal membrane integrity and fibrosis.
- Enables continuity from mechanistic discovery to preclinical validation of candidate therapeutics.
- Supports risk-adjusted advancement by providing reliable data on intervention efficacy in a disease-relevant system.
- Facilitates biomarker discovery and validation for translational research in peritoneal dialysis complications.
Pipeline & Workflow Integration
This improved murine PD model integrates into the discovery-to-preclinical continuum, supporting target validation, lead identification, and translational research on peritoneal membrane failure.
- Discovery Biology: Enables rigorous hypothesis testing and mechanistic de-risking for peritoneal fibrosis pathways.
- Screening: Provides a reproducible in vivo assay platform for quantitative evaluation of candidate interventions.
- Analytics: Delivers measurable outputs such as ECM deposition, fibrosis intensity, and vascularity for comparative analysis.
- Translational Research: Bridges discovery findings to preclinical validation in a clinically relevant context.
- Enterprise Reuse: Establishes a standardized, durable model for ongoing and future R&D programs targeting peritoneal complications.
Operational & Enterprise Impact
- Scientific Value: Increases predictive confidence and reduces mechanistic ambiguity in peritoneal fibrosis research.
- Operational Value: Enhances model standardization, reproducibility, and scalability for multi-study use.
- Strategic Value: Supports informed go/no-go decisions and capital-efficient portfolio management.
- Portfolio Impact: Enables risk-adjusted prioritization and advancement of therapeutic candidates targeting peritoneal membrane failure.
Implementation Considerations
- Requires surgical expertise in murine catheter implantation and perioperative care.
- Needs access to imaging, histological, and quantitative analytical infrastructure for endpoint assessment.
- Demands cross-team standardization of surgical and analytical protocols to ensure reproducibility.
- May require adaptation for different mouse strains or disease models as supported by study scope.
- Technical learning curve and procedural nuances must be addressed for consistent model generation.
Biopharma FAQs
Chapters in this video
0:04
Introduction
1:18
Skin Preparation
1:42
Measure the Catheter Length and Mark the Insertion Point Within the Abdomen and the Tube Tract Over the Prepared Skin
2:12
Customize the Peritoneal Catheter Reservoir Section and Place the Instillation Port
2:52
Make the Catheter Tip Insertion Site Incision and Fix the Tip Inside the Abdominal Cavity and Fluid Injections
4:15
Peritoneal Biopsy
4:45
Results: Improved Method for Implanting a Peritoneal Dialysis Catheter in a Murine Model
5:47
Conclusion
