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通过逆行灌注酶消化法分离单个小鼠心肌细胞的简易方法。

Simple Method to Isolate Individual Mouse Cardiomyocytes Using Enzymatic Digestion via Retrograde Perfusion.

作者信息

Hintz Madyson O, Zellhuber-McMillan Ute, Jones Peter P

机构信息

Department of Physiology and HeartOtago, School of Biomedical Sciences, University of Otago, Otago, New Zealand.

出版信息

Methods Mol Biol. 2025;2894:7-20. doi: 10.1007/978-1-0716-4342-6_2.

Abstract

Isolated single cardiomyocytes are commonly used to understand fundamental cardiac cell physiology and cell signaling at the molecular level. The protocol to isolate cardiomyocytes is under continual optimization between laboratories, despite being conducted for many years. The delicate nature of these cells coinciding with laboratory equipment variations can make the consistency of isolation challenging. The protocol described here presents a simple method to routinely isolate high-quality cardiomyocytes using Langendorff perfusion. This method provides cells well suited for analysis of physiological cellular calcium handling and spontaneous calcium release.

摘要

分离的单个心肌细胞常用于在分子水平上理解基本的心脏细胞生理学和细胞信号传导。尽管心肌细胞分离方案已经实施多年,但各实验室仍在不断优化该方案。这些细胞的脆弱性与实验室设备的差异相结合,可能使分离的一致性具有挑战性。本文所述的方案提出了一种使用Langendorff灌注常规分离高质量心肌细胞的简单方法。该方法提供的细胞非常适合用于分析生理性细胞钙处理和自发性钙释放。

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