Optimized Langendorff perfusion system for cardiomyocyte isolation in adult mouse heart.
Haotong LiChungeng LiuMinghui BaoWeijing LiuNie YuHong LianShengshou HuPublished in: Journal of cellular and molecular medicine (2020)
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.
Keyphrases
- single cell
- contrast enhanced
- cardiovascular disease
- high glucose
- heart failure
- angiotensin ii
- magnetic resonance imaging
- computed tomography
- randomized controlled trial
- type diabetes
- rna seq
- endothelial cells
- metabolic syndrome
- magnetic resonance
- high throughput
- cardiovascular risk factors
- young adults
- loop mediated isothermal amplification