Key Points
- To establish and validate a fast, efficient technique for isolating viable cardiac myocytes and fibroblasts from postnatal mice at any developmental age, specifically closing the gap between three days and three weeks post-birth.
- Perfused whole hearts through the left ventricle at a high flow rate using a simplified syringe-pump apparatus.
- Digested cardiac tissue using an enzymatic solution containing trypsin and collagenase in a Krebs-based buffer.
- Successfully isolated fresh, intact cardiomyocytes from postnatal mice regardless of age, overcoming the conventional 3-day to 3-week developmental barrier.
- Achieved a high digestion throughput of 10 to 15 hearts per hour, markedly reducing cell isolation time compared to traditional protocols.
Structured PICO
PPopulationPostnatal mouse hearts (neonatal and adult mice)
IInterventionWhole heart perfusion of a trypsin-collagenase Krebs-based buffer through the left ventricle at a high flow rate using a syringe pump
OOutcomeSuccessful isolation of viable cardiomyocytes and fibroblasts
A novel, rapid syringe-pump-based perfusion method enables efficient isolation of cardiomyocytes from postnatal mice of any age, overcoming limitations of traditional Langendorff and neonatal protocols.