Journal
JOURNAL OF CELLULAR AND MOLECULAR MEDICINE
Volume 24, Issue 24, Pages 14619-14625Publisher
WILEY
DOI: 10.1111/jcmm.15773
Keywords
adult mouse heart; cardiomyocyte isolation; Langendorff perfusion system
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Funding
- National Natural Science Foundation of China [81900343]
- Beijing Natural Science Foundation [7182140]
- Chinese Academy of Medical Sciences Innovation Fund for Medical Sciences [2016-I2M-1-015]
- National Key Research and Development Project of China [2019YFA0801500]
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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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