Key result
Gravity-based Langendorff perfusion increases viable cardiomyocyte yield to ~83% compared with peristaltic pumps.
Why the study?
Traditional Langendorff perfusion systems are difficult to master, and cardiomyocyte viability and purity frequently fail to meet sequencing requirements due to complicated devices and manipulation processes.
Absolute Event Rate: 83.1% vs 70.97%
p-value: p=<0.05
An optimized Langendorff perfusion system provides a simplified, standardized method for isolating high-viability and high-purity cardiomyocytes from adult mouse hearts for downstream applications like single-cell sequencing.
May improve rodent cardiomyocyte yields for sequencing; leaves open translation to human or clinical applications.
With the rapid development of single-cell sequencing technology, the Langendorff perfusion system has emerged as a common approach to decompose cardiac tissue and obtain living cardiomyocytes to study cardiovascular disease with the mechanism of cardiomyocyte biology. However, the traditional Langendorff perfusion system is difficult to master, and further, the viability and purity of cardiomyocytes are frequently unable to meet sequencing requirements due to complicated devices and manipulate processes. Here, we provide an optimized Langendorff perfusion system with a simplified and standardized operating protocol which utilizes gravity as the perfusion pressure, includes a novel method for bubbles removing and standardizes the criteria for termination of digestion. We obtained stable cardiomyocyte with high viability and purity after multiple natural gravity sedimentation. The combination of the optimized Langendorff perfusion system and the multiple natural gravity sedimentation provides a stable system for isolating adult mouse heart, which will provide higher-quality cardiomyocytes for further experiments.
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Li et al. (2020) studied Cardiomyocyte isolation. Optimized Langendorff perfusion system with gravity pressure vs. Classical Langendorff method using peristaltic pumps was evaluated on Rate of viable rod-shaped cardiomyocytes (p=<0.05). The optimized Langendorff perfusion system utilizing gravity as perfusion pressure significantly improved the rate of viable rod-shaped cardiomyocytes to 83.1% compared to 71.0% with the classical peristaltic pump method.
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