A simple antegrade perfusion method for isolating viable single cardiomyocytes from neonatal to aged mice

Area of Science:

• Cardiology
• Cell Biology
• Biomedical Engineering

Background:

• Isolating viable single heart cells is crucial for studying cardiac function and disease.
• Traditional Langendorff retrograde perfusion methods have limitations in efficiency and applicability across different age groups.
• A need exists for a simplified and adaptable method for obtaining high-quality cardiac myocytes.

Purpose of the Study:

• To develop and validate a simple, reproducible antegrade perfusion method for isolating single viable mouse heart cells.
• To establish protocols suitable for mice of various ages, from neonatal to aged.
• To compare the quality and characteristics of cells isolated by antegrade perfusion with those from traditional retrograde methods.

Main Methods:

• Antegrade perfusion was performed by injecting perfusate into the left ventricle of excised mouse hearts with a clamped aorta.
• Procedures were conducted under microscopy on a prewarmed mat to monitor injection and perfusion.
• Adjustments in needle size, enzyme solution, and flow rate allowed application across different ages.

Main Results:

• The antegrade perfusion method successfully isolated single viable ventricular and atrial myocytes from mice of all ages.
• Isolated cells exhibited morphological and electrophysiological properties comparable to those obtained via retrograde perfusion.
• Interstitial nonmyocytes, including cardiac progenitor cells, were also isolated from the supernatant.

Conclusions:

• The developed antegrade perfusion method is a simple and effective technique for high-quality isolation of mouse heart cells.
• This method is adaptable for use with mouse hearts from neonatal to aged individuals.
• The antegrade approach offers a viable alternative to traditional methods for cardiac cell isolation.