Li Haotong, Liu Chungeng, Bao Minghui, Liu Weijing, Nie Yu, Lian Hong, Hu Shengshou
State Key Laboratory of Cardiovascular Disease, National Center for Cardiovascular Disease, Chinese Academy of Medical Sciences and Peking Union Medical College, Fuwai Hospital, Beijing, China.
Department of Cardiovascular Surgery, Tongji Medical College, Union Hospital, Huazhong University of Science and Technology, Wuhan, China.
J Cell Mol Med. 2020 Dec;24(24):14619-14625. doi: 10.1111/jcmm.15773. Epub 2020 Nov 4.
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.
随着单细胞测序技术的快速发展，Langendorff灌注系统已成为一种常用的方法，用于分解心脏组织并获取活的心肌细胞，以研究心血管疾病的心肌细胞生物学机制。然而，传统的Langendorff灌注系统难以掌握，而且由于设备复杂和操作过程，心肌细胞的活力和纯度常常无法满足测序要求。在此，我们提供了一种优化的Langendorff灌注系统，其具有简化和标准化的操作方案，利用重力作为灌注压力，包括一种新颖的气泡去除方法，并规范了消化终止标准。经过多次自然重力沉降后，我们获得了具有高活力和纯度的稳定心肌细胞。优化的Langendorff灌注系统与多次自然重力沉降相结合，为分离成年小鼠心脏提供了一个稳定的系统，这将为进一步的实验提供更高质量的心肌细胞。
