Large Volume Blood Collection from Swine for In Vitro Applications: Use of Intracardiac Cannulation and a Vacuum Pump as a Terminal Procedure
July 25th, 2025
Here we present a method to collect a large volume of blood from swine using percutaneous intracardiac cannulation connected to a vacuum pump, as a terminal procedure.
This procedure allow us the opportunity to collect a large volume of blood that maintains cell integrity to be used in master seeds virus development. There is variation in the pressure applied by personnel that may compromise, damage, disrupt, or impair the cells. Our efficient technique provides excellent cell viability and constant vacuum suction to maximize collection volume.
Large volume blood draws can be performed manually, but there may be variation in the pressure applied. Our efficient technique provides excellent cell viability and constant vacuum suction to maximize collection volume and quality. To begin, assemble the tubing to the stainless steel straws and connect the hose end to the bubble tubing.
Attach the tubing assembly to the roller bottle using a rubber stopper fitted with two uneven metal straws and a glass bottle. Aseptically instill 10 milliliters of sodium heparin solution through the suction tubing and catheter stylet to prevent clotting. Then add 50 milliliters of sodium heparin solution at the same concentration into each roller bottle.
Confirm the vacuum pump is set to no more than seven pounds of draw. Next, place an anesthetized pig on a lift table. Clip the thoracic hair over the heart near the fourth and fifth intercostal space just caudal to the point of the elbow.
If available, use ultrasound to identify the left ventricle. If ultrasound is not available, locate the left ventricle by auscultating and palpating near the fourth or fifth intercostal space medial to the point of maximal impulse. Next, aseptically scrub the area with four by four inch gauze sponges soaked in 2%chlorhexidine scrub, followed by gauze sponges soaked in 70%isopropyl alcohol.
Scrub in a circular manner, starting at the point of injection and rotating outwards. Repeat this process at least three times, alternating between the two disinfectants. Verify that the animal remains under surgical anesthesia by checking for absent palpebral reflex, loose jaw tone, and absence of pedal or withdrawal reflex.
Now remove the stylet from the intravenous catheter. Position the stylet over the left ventricle and insert it perpendicularly through the skin into the ventricle. When blood begins to pulse out of the stylet, connect it to the bubble tubing hose end attached to the roller bottle and activate the vacuum pump.
Confirm a steady stream of blood flowing into the roller bottle. As blood is pulled into the roller bottle, gently swirl the roller bottle to mix the blood with the anticoagulant. When a liter of blood has been collected, use hemostats to clamp the bubble tubing near the roller bottle.
Then disconnect the rubber stopper and attach it to the next roller bottle. Release the hemostat to resume flow and continue swirling gently. Continue collecting blood in this manner until the desired volume is reached.
Clamp the tubing and turn off the pump to end the collection. Place the cleaned rubber stoppers, straws, hose end connectors, and hemostats into a new autoclave pouch and sterilize before the next use.
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Overview
This article presents a method for collecting a large volume of blood from swine using percutaneous intracardiac cannulation connected to a vacuum pump. The technique aims to maintain cell integrity for master seed virus development.
Key Study Components
Area of Science
- Neuroscience
- Biology
- Veterinary Medicine
Background
- Blood collection is critical for various biological and medical research.
- Maintaining cell integrity during collection is essential for downstream applications.
- Traditional methods may lead to variability in pressure applied during blood draws.
- Efficient techniques are needed to enhance cell viability and collection volume.
Purpose of Study
- To develop a reliable method for large volume blood collection from swine.
- To ensure high cell viability during the collection process.
- To minimize variability associated with manual blood draws.
Methods Used
- Percutaneous intracardiac cannulation.
- Connection to a vacuum pump for constant suction.
- Assembly of tubing to stainless steel straws.
- Utilization of bubble tubing for enhanced collection efficiency.
Main Results
- The technique provides excellent cell viability.
- Constant vacuum suction maximizes blood collection volume.
- Reduces the risk of cell damage during the collection process.
- Demonstrates a significant improvement over manual collection methods.
Conclusions
- The developed method is efficient for large volume blood draws.
- Maintaining cell integrity is crucial for subsequent research applications.
- This technique can enhance the quality of biological samples collected from swine.
Frequently Asked Questions
Executive Industry Relevance
Reliable access to large volumes of high-integrity swine blood is critical for downstream in vitro applications such as primary cell culture and master seed virus development. This terminal intracardiac cannulation method with vacuum-assisted collection addresses the need for scalable, reproducible sample acquisition while minimizing cell lysis and variability. The approach supports enterprise R&D by enabling consistent biological inputs for assay development and translational workflows.
Strategic Applications in Biopharma R&D
Early Discovery & Target Validation
- Enables acquisition of high-viability blood cells for mechanistic studies and target interrogation.
- Reduces biological variability, supporting functional validation of cellular models.
- Facilitates predictive confidence in downstream cell-based assays by preserving sample quality.
Screening & Assay Development
- Provides standardized, large-volume blood samples for assay optimization and reproducibility.
- Supports preparation of validated biological systems for compound screening and evaluation.
- Minimizes cell damage, ensuring reliable quantitative outputs in primary cell-based assays.
Translational & Preclinical Research
- Enables continuity from sample collection to preclinical model development when high-integrity blood is required.
- Supports alignment with translational biomarker studies by maintaining cellular integrity.
- Reduces risk of confounding variables in disease-relevant systems due to sample inconsistency.
Pipeline & Workflow Integration
This method integrates at the interface of biological sample acquisition and in vitro assay development, supporting workflows from early discovery through preclinical research.
- Discovery Biology: Provides high-quality blood for hypothesis testing and pathway analysis.
- Screening: Delivers reproducible, scalable samples for assay readiness and standardization.
- Analytics: Enables quantitative assessment of cell viability and sample integrity across conditions.
- Translational Research: Maintains sample quality for biomarker and preclinical studies when supported by downstream needs.
- Enterprise Reuse: Establishes a standardized protocol for repeated, high-volume blood collection across programs.
Operational & Enterprise Impact
- Scientific Value: Enhances predictive confidence and reduces mechanistic ambiguity in cell-based research.
- Operational Value: Improves standardization, reproducibility, and scalability of blood collection workflows.
- Strategic Value: Supports better go/no-go decisions by ensuring reliable biological inputs for R&D pipelines.
- Portfolio Impact: Enables risk-adjusted prioritization by reducing late-stage biological variability.
Implementation Considerations
- Requires personnel trained in aseptic technique and terminal anesthesia procedures.
- Needs access to vacuum pumps, sterile tubing assemblies, and anticoagulant preparation.
- Demands cross-team standardization to ensure consistent sample quality across studies.
- Adaptation to other large animal models may require protocol modification and validation.
- Terminal nature of the procedure limits reuse per animal and necessitates ethical oversight.
