Slicing and Culturing Pig Hearts under Physiological Conditions
March 20th, 2020
17.6K views
This protocol describes how to slice and culture heart tissue under physiological conditions for 6 days. This culture system could be used as a platform for testing the efficacy of novel heart failure therapeutics as well as reliable testing of acute cardiotoxicity in a 3D heart model.
Overview
This protocol describes the slicing and culturing of heart tissue sections for six days under physiological conditions. This system can be utilized for testing the efficacy of heart failure therapies and assessing acute cardiac toxicity.
Key Study Components
Area of Science
- Cardiac research
- Tissue culture
- Pharmacology
Background
- Heart failure is a significant health issue requiring effective therapies.
- Acute cardiac toxicity testing is essential for drug development.
- Current models may not accurately predict human responses.
- This protocol aims to bridge the gap between pre-clinical and clinical testing.
Purpose of Study
- To provide a reliable method for heart tissue culture.
- To evaluate the efficacy of novel heart failure therapeutics.
- To establish a predictive model for acute cardiac toxicity testing.
Methods Used
- Use of a vibratome for slicing heart tissue.
- Preparation of cardioplegic solution for tissue preservation.
- Isolation of the left ventricle for culturing.
- Culture maintenance under physiological conditions for six days.
Main Results
- Successful culture of heart tissue sections for six days.
- Potential for testing heart failure therapies demonstrated.
- Acute cardiotoxicity testing model established.
- Protocol provides a reproducible method for future studies.
Conclusions
- The developed culture system is a valuable tool for cardiac research.
- It enhances the predictive accuracy of drug testing.
- This method could lead to improved therapeutic strategies for heart failure.
Frequently Asked Questions
What type of heart tissue is used in this protocol?
Pig heart tissue is utilized for slicing and culturing.
How long is the heart tissue cultured?
The heart tissue is cultured for six days under physiological conditions.
What is the purpose of using a vibratome?
A vibratome is used to slice the heart tissue into precise sections for culture.
What is the significance of the cardioplegic solution?
The cardioplegic solution helps preserve the heart tissue during the slicing process.
How does this protocol improve drug testing?
It provides a more accurate predictive model for assessing drug efficacy and toxicity.
Executive Industry Relevance
Current in vitro and in vivo models poorly predict human cardiac liabilities, leading to costly late-stage failures in drug development. This protocol provides a biomimetic heart slice culture system that preserves viability and function for six days under physiological conditions, enabling more accurate assessment of acute cardiotoxicity and therapeutic efficacy. By using multicellular adult heart tissue from pig (and translatable to human) sources, the model addresses a critical gap in preclinical safety testing and supports better go/no-go decisions in cardiovascular drug discovery.
Strategic Applications in Biopharma R&D
Early Discovery & Target Validation
- Scientific Value: Enables interrogation of therapeutic hypotheses in multicellular adult heart tissue that reflects human myocardium physiology.
- Operational Value: Supports functional target validation by measuring contractile force and responses to beta-adrenergic stimulation over six days in culture.
Screening & Assay Development
- Scientific Value: Provides a standardized, reproducible platform for preparing validated heart slices for downstream compound evaluation.
- Operational Value: Supports assay readiness through consistent tissue slicing, mounting, and culture under electrical stimulation and oxygenation.
Translational & Preclinical Research
- Scientific Value: Uses adult cardiomyocytes with physiological properties distinct from immature hiPSC-CMs, improving translational relevance.
- Operational Value: Enables continuity from discovery through preclinical validation by maintaining structural and functional biomarkers for up to six days.
Pipeline & Workflow Integration
The method fits within the discovery continuum from early target validation to preclinical assessment, supporting hypothesis testing and lead identification through functional readouts.
- Discovery Biology: Supports pathway clarification and biological de-risking by enabling testing in intact multicellular tissue under near-physiological conditions.
- Screening: Delivers assay readiness and reproducibility via standardized slicing, mounting, and culture procedures with electrical pacing.
- Analytics: Provides quantitative outputs including MTT viability, contractile force, calcium transient measurements, and metabolic assessments for comparative condition analysis.
- Translational Research: Connects to preclinical continuity through preservation of adult cardiomyocyte function and structural integrity over six days.
- Enterprise Reuse: Establishes a reusable platform applicable to pig, human, dog, sheep, or cat heart slices for cardiotoxicity and efficacy testing across programs.
Operational & Enterprise Impact
- Scientific Value: Increases predictive confidence by modeling human-like multicellular heart tissue responses to drugs and gene therapies.
- Operational Value: Ensures reproducibility through standardized vibratome slicing, tissue mounting, and culture under electrical stimulation (1.2 Hz, 10 V) and oxygenation.
- Strategic Value: Reduces late-stage biological risk by improving early detection of cardiotoxicity, supporting capital-efficient advancement decisions.
- Portfolio Impact: Enables risk-adjusted prioritization of compounds based on functional heart slice responses, improving go/no-go criteria in cardiovascular portfolios.
Implementation Considerations
- Requires expertise in tissue handling, vibratome operation, and sterile technique for heart dissection and slicing.
- Depends on specialized equipment including vibratome, cell culture electrical stimulator, oxygenation setup, and incubators with 5% CO2 at 37°C.
- Necessitates cross-team standardization of tissue preparation, mounting on polyurethane belts, and media change protocols to ensure slice viability.
- Involves adaptation considerations for different species (pig, human, dog, sheep, cat) and tissue block sizing (1–2 cm³) to maintain slicing quality.
- Includes practical limitations such as the need for daily media changes, stimulation plate maintenance, and avoidance of graphite particle contamination during electrical stimulation.
Biopharma FAQs
Why is null hypothesis testing important for validating cardiac targets in heart slice models?
Null hypothesis testing helps determine whether observed changes in contractile force or calcium transients after compound exposure are statistically significant, supporting objective target validation in multicellular tissue.
How does isolating independent variables like electrical stimulation frequency improve target validation in heart slice cultures?
Controlling variables such as 1.2 Hz electrical stimulation ensures that observed functional responses are attributable to the test compound rather than culture artifacts, improving target validation reliability.
What quantitative dependent variable measurements enable assessment of drug effects in cultured heart slices?
Measurements such as MTT viability, contractile force, calcium transient amplitude, and metabolic readouts provide quantitative endpoints to evaluate drug-induced changes in heart slice function.
Why are replication requirements critical for ensuring cross-functional collaboration in heart slice-based toxicity testing?
Replication across slices and experiments ensures data consistency, enabling toxicology, pharmacology, and drug safety teams to rely on reproducible results for go/no-go decisions.
What statistical analysis capabilities are required before implementing heart slice models in drug screening workflows?
Teams require the ability to perform t-tests or ANOVA on functional readouts like contractility or viability to assess statistical significance of drug effects across treatment and control groups.
