Methods in cardiomyocyte isolation, culture, and gene transfer |
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Authors: | Louch William E Sheehan Katherine A Wolska Beata M |
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Affiliation: | a Institute for Experimental Medical Research, Oslo University Hospital Ullevaal, Oslo, Norwayb Centre for Heart Failure Research, Faculty of Medicine, University of Oslo, Oslo, Norwayc Department of Physiology and Biophysics, University of Illinois at Chicago, Chicago, IL 60612, USAd Department of Medicine, Section of Cardiology, Center for Cardiovascular Research, University of Illinois at Chicago, Chicago, IL 60612, USA |
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Abstract: | Since techniques for cardiomyocyte isolation were first developed 35 years ago, experiments on single myocytes have yielded great insight into their cellular and sub-cellular physiology. These studies have employed a broad range of techniques including electrophysiology, calcium imaging, cell mechanics, immunohistochemistry and protein biochemistry. More recently, techniques for cardiomyocyte culture have gained additional importance with the advent of gene transfer technology. While such studies require a high quality cardiomyocyte population, successful cell isolation and maintenance during culture remain challenging. In this review, we describe methods for the isolation of adult and neonatal ventricular myocytes from rat and mouse heart. This discussion outlines general principles for the beginner, but also provides detailed specific protocols and advice for common caveats. We additionally review methods for short-term myocyte culture, with particular attention given to the importance of substrate and media selection, and describe time-dependent alterations in myocyte physiology that should be anticipated. Gene transfer techniques for neonatal and adult cardiomyocytes are also reviewed, including methods for transfection (liposome, electroporation) and viral-based gene delivery. |
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Keywords: | Methods Cardiomyocyte Isolation Cell culture Gene transfer Transfection Transduction |
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