With or Without Langendorff
A New Method for Adult Myocyte Isolation to Be Tested With Time
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To study molecular and cellular mechanisms for normal cardiac function and cardiac pathology, high-quality isolated adult cardiomyocytes are essential. Isolated cardiomyocytes can be used directly for cellular function studies such as measuring myocyte contraction, cellular electrophysiology, calcium imaging, myofilament calcium sensitivity, cytoskeletal function, drug effects, and metabolism. Isolated myocytes can also be cultured for signaling studies with less contamination from nonmyocytes. In addition, the isolated cells are also essential for studying the morphology and structure of cardiomyocytes. Furthermore, the function of nonmyocytes including cardiac fibroblasts, smooth muscle cells, infiltrated inflammatory cells, and endothelial cells in the heart is also an important topic in cardiac biology, and there is an increasing appreciation of the structure and function of these cells in cardiac pathology. Therefore, the development of techniques for heart cell isolation and harvest dates back to ≈40 years ago. In the current issue of Circulation Research, Ackers-Johnson et al1 describe a simple approach for the isolation and culture of adult mouse cardiac myocytes and fibroblasts without the traditional Langendorff perfusion system.
Article, see p 909
Although techniques for isolation of adult ventricular myocytes from many species including rat,2 rabbit,3 and canine4 have been available for ≈40 years, there remains no clear consensus on the optimal protocol to isolate and culture viable adult cardiac myocytes. However, at their core, these protocols generally all rely on digestion of the heart with an enzymatic solution.5 All myocyte isolation techniques can be essentially categorized into either chunk (a small piece of tissue) digestion in an enzymatic solution or coronary artery perfusion with enzymatic solution. The chunk digestion technique basically allows the enzymes (collagenases supplemented with other proteases) to chew the extracellular matrix directly and dissociate cardiomyocytes and nonmyocytes from small pieces (normally <1 mm in diameter) of cardiac …