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Merck
CN

Acute Isolation of Cells from Murine Sino-atrial Node.

Bio-protocol (2020-01-05)
Qadeer Aziz, Muriel Nobles, Andrew Tinker
摘要

The cardiac conduction system allows the synchronized propagation of electrical activity through heart muscle. This is initiated by the spontaneous activity of the specialized pacemaker cells of the sino-atrial node (SAN). The SAN region underlies automaticity in mammals and therefore has a crucial role in the pathogenesis of cardiac disorders such as arrhythmia. Isolation of SAN tissue and SAN cells is critical to advance our understanding of SAN structure and function in health and disease. Initially, isolation of SAN tissue and SAN cells was carried out in the rabbit owing to its larger size and similar electrical properties to human. This protocol was optimized by Mangoni and Nargeot (2001) for use in mice to take advantage of advancements in transgenic models. Here, we provide a step-by-step guide to dissecting the SAN tissue and isolating pacemaker cardiomyocytes from mouse hearts using an enzyme digestion approach.

材料
Product Number
品牌
产品描述

Sigma-Aldrich
层粘连蛋白 来源于 Engelbreth-Holm-Swarm 小鼠肉瘤基底膜, 1-2 mg/mL in Tris-buffered saline, 0.2 μm filtered, BioReagent, suitable for cell culture
Sigma-Aldrich
磷酸钾 一元, ACS reagent, ≥99.0%
Sigma-Aldrich
蛋白酶 来源于灰色链霉菌, Type XIV, ≥3.5 units/mg solid, powder
Sigma-Aldrich
氯化钠, BioXtra, ≥99.5% (AT)
Sigma-Aldrich
L-谷氨酸 钾盐 一水合物, ≥99% (HPLC), powder
Sigma-Aldrich
D -(+)-葡萄糖, ACS reagent
Sigma-Aldrich
氯化钾, BioXtra, ≥99.0%