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Isolation and Functional Characterization of Human Ventricular Cardiomyocytes from Fresh Surgical Samples
Published on: April 21, 2014
Isolation and functional characterization of human ventricular cardiomyocytes from fresh surgical samples
Raffaele Coppini1, Cecila Ferrantini2, Alessandro Aiazzi2
1Department NeuroFarBa, Division of Pharmacology, University of Florence; raffaele.coppini@unifi.it.
Insights
Researchers developed a new protocol to isolate viable human cardiomyocytes from heart surgery samples. This method enables crucial studies on heart disease mechanisms and potential new treatments.
Area of Science:
- Cardiology
- Cell Biology
- Physiology
Background:
- Diseased heart cells undergo remodeling affecting function and increasing arrhythmia risk.
- Current understanding of myocyte dysfunction in heart disease primarily relies on animal models.
Purpose of the Study:
- To describe and validate a protocol for isolating viable human ventricular cardiomyocytes.
- To enable direct study of human cardiomyocyte function in cardiac diseases.
Main Methods:
- Isolation of viable cardiomyocytes from small surgical samples of human ventricular myocardium.
- Demonstration of feasibility using electrophysiological and intracellular calcium measurements.
Main Results:
- Successful isolation of viable human ventricular cardiomyocytes.
- Demonstrated feasibility of single-cell measurements (electrophysiology, calcium handling) in isolated human cells.
Conclusions:
- The protocol provides a valuable tool for investigating the cellular basis of human heart disease.
- This method facilitates the identification of therapeutic targets and testing of novel compounds on human cardiomyocytes.
Abstract:
Cardiomyocytes from diseased hearts are subjected to complex remodeling processes involving changes in cell structure, excitation contraction coupling and membrane ion currents. Those changes are likely to be responsible for the increased arrhythmogenic risk and the contractile alterations leading to systolic and diastolic dysfunction in cardiac patients. However, most information on the alterations of myocyte function in cardiac diseases has come from animal models. Here we describe and validate a protocol to isolate viable myocytes from small surgical samples of ventricular myocardium from patients undergoing cardiac surgery operations. The protocol is described in detail. Electrophysiological and intracellular calcium measurements are reported to demonstrate the feasibility of a number of single cell measurements in human ventricular cardiomyocytes obtained with this method. The protocol reported here can be useful for future investigations of the cellular and molecular basis of functional alterations of the human heart in the presence of different cardiac diseases. Further, this method can be used to identify novel therapeutic targets at cellular level and to test the effectiveness of new compounds on human cardiomyocytes, with direct translational value.
