Optimized Langendorff perfusion system for cardiomyocyte isolation in adult mouse heart.
Optimized Langendorff perfusion system for cardiomyocyte isolation in adult mouse heart.
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用于成年小鼠心脏心肌细胞分离的优化 Langendorff 灌注系统
DOI：
10.1111/jcmm.15773
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发表时间：
2020-12
影响因子：
5.3
通讯作者：
Hu S
中科院分区：
文献类型：
--
作者：
Li H;Liu C;Bao M;Liu W;Nie Y;Lian H;Hu S
随着单细胞测序技术的快速发展，Langendorff灌注系统已成为分解心脏组织并获得活心肌细胞的常用方法，以心肌细胞生物学机制研究心血管疾病。然而，传统的Langendorff灌注系统难以掌握，并且由于复杂的设备和操作过程，心肌细胞的活力和纯度常常无法满足测序要求。在这里，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，该操作方案利用重力作为灌注压力，包括用于气泡去除的新方法，并简化了消化终止的标准。经过多次自然重力沉降，获得了稳定的高活力、高纯度的心肌细胞。优化的Langendorff灌流系统和多次自然重力沉降的组合为分离成年小鼠心脏提供了稳定的系统，这将为进一步的实验提供更高质量的心肌细胞。
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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影响因子：
20.1
作者：
Ackers-Johnson M;Li PY;Holmes AP;O'Brien SM;Pavlovic D;Foo RS
通讯作者：
Foo RS
用于心脏再生和修复的细胞外基质生物材料
影响因子：
4.6
作者：
Li, Haotong;Bao, Minghui;Nie, Yu
通讯作者：
Nie, Yu
影响因子：
5.5
作者：
Nguyen QH;Pervolarakis N;Nee K;Kessenbrock K
通讯作者：
Kessenbrock K
影响因子：
37.8
作者：
Quaife-Ryan GA;Sim CB;Ziemann M;Kaspi A;Rafehi H;Ramialison M;El-Osta A;Hudson JE;Porrello ER
通讯作者：
Porrello ER
影响因子：
1.2
作者：
Judd, Justin;Lovas, Jonathan;Huang, Guo N.
通讯作者：
Huang, Guo N.