Blackwood Erik A, Bilal Alina S, Azizi Khalid, Sarakki Anup, Glembotski Christopher C
San Diego State University Heart Institute and the Department of Biology, San Diego State University.
San Diego State University Heart Institute and the Department of Biology, San Diego State University;
J Vis Exp. 2020 Jun 14(160). doi: 10.3791/61224.
The isolation and culturing of cardiac myocytes from mice has been essential for furthering the understanding of cardiac physiology and pathophysiology. While isolating myocytes from neonatal mouse hearts is relatively straightforward, myocytes from the adult murine heart are preferred. This is because compared to neonatal cells, adult myocytes more accurately recapitulate cell function as it occurs in the adult heart in vivo. However, it is technically difficult to isolate adult mouse cardiac myocytes in the necessary quantities and viability, which contributes to an experimental impasse. Furthermore, published procedures are specific for the isolation of either atrial or ventricular myocytes at the expense of atrial and ventricular non-myocyte cells. Described here is a detailed method for isolating both atrial and ventricular cardiac myocytes, along with atrial and ventricular non-myocytes, simultaneously from a single mouse heart. Also provided are the details for optimal cell-specific culturing methods, which enhance cell viability and function. This protocol aims not only to expedite the process of adult murine cardiac cell isolation, but also to increase the yield and viability of cells for investigations of atrial and ventricular cardiac cells.
从小鼠中分离和培养心肌细胞对于深入了解心脏生理学和病理生理学至关重要。虽然从新生小鼠心脏中分离心肌细胞相对简单，但成年小鼠心脏的心肌细胞更受青睐。这是因为与新生细胞相比，成年心肌细胞能更准确地重现成年心脏在体内发生的细胞功能。然而，从成年小鼠心脏中分离出足够数量且具有活力的心肌细胞在技术上具有挑战性，这导致了实验僵局。此外，已发表的方法专门用于分离心房或心室肌细胞，却牺牲了心房和心室的非心肌细胞。本文描述了一种从单个小鼠心脏同时分离心房和心室心肌细胞以及心房和心室非心肌细胞的详细方法。还提供了优化的细胞特异性培养方法的细节，这些方法可提高细胞活力和功能。该方案不仅旨在加快成年小鼠心脏细胞分离的过程，还旨在提高用于心房和心室心脏细胞研究的细胞产量和活力。
