Institute for Experimental Medical Research, Oslo University Hospital Ullevaal, Oslo, Norway.
J Mol Cell Cardiol. 2011 Sep;51(3):288-98. doi: 10.1016/j.yjmcc.2011.06.012. Epub 2011 Jun 24.
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.
自 35 年前首次开发出心肌细胞分离技术以来，对单个心肌细胞的实验已经深入了解了它们的细胞和亚细胞生理学。这些研究采用了广泛的技术，包括电生理学、钙成像、细胞力学、免疫组织化学和蛋白质生物化学。最近，随着基因转移技术的出现，心肌细胞培养技术获得了更多的重视。虽然这些研究需要高质量的心肌细胞群体，但在培养过程中成功的细胞分离和维持仍然具有挑战性。在这篇综述中，我们描述了从大鼠和小鼠心脏中分离成年和新生心室心肌细胞的方法。这部分内容概述了初学者的一般原则，但也为常见注意事项提供了详细的具体方案和建议。我们还回顾了短期心肌细胞培养的方法，特别关注基质和培养基选择的重要性，并描述了预期的心肌细胞生理学的时间依赖性变化。还回顾了成年和新生心肌细胞的基因转移技术，包括转染（脂质体、电穿孔）和基于病毒的基因传递方法。
