PDF(Pubmed)
Abstract：
Living heart slices have recently emerged as a powerful experimental model for fundamental cardiac research. By retaining the structure and function of the native myocardium while maintaining the simplicity of cell culture models, heart slices can be easily employed in electrophysiological, pharmacological, biochemical, and structural investigations. One single heart yields many slices (>20 slices for rodents, >100 slices for porcine or human hearts), however due to the low throughput of most assays and rapid slice degeneration within 24 h of preparation, many slices remain unused and are discarded at the end of the preparation day. Here we present a novel method to extend viability and functionality of living heart slices, enabling their use in experiments over several consecutive days following preparation. By combining hypothermic conditions with inhibition of myosin II ATPase using 2,3-butanedione monoxime (BDM), slices prepared from the left ventricle of porcine hearts remain viable and exhibit preserved contractile function and morphology for up to 6 days. Electrophysiological function was also confirmed over the 6 days by extracellular field potentials recordings. This simple method not only maximizes the use of slices prepared from one single heart, thus reducing the number of animals required, but also increases data reproducibility by allowing multiple electrophysiological, pharmacological, biochemical, and structural studies to be performed from the same heart.
摘要：
最近，活体心脏切片已成为基础心脏研究的强大实验模型。通过保留天然心肌的结构和功能，同时保持细胞培养模型的简单性，心脏切片可以很容易地用于电生理，药理学,生物化学，和结构调查。一个心脏产生许多切片（啮齿动物>20切片，>100片用于猪或人的心脏），然而，由于大多数分析的低通量和24小时内制备的快速切片变性，许多切片仍未使用，并在准备日结束时丢弃。在这里，我们提出了一种新的方法来扩展活心脏切片的活力和功能，使它们能够在制备后的连续几天内用于实验。通过将低温条件与使用2,3-丁二酮单肟（BDM）抑制肌球蛋白IIATPase相结合，从猪心脏的左心室制备的切片保持存活并且表现出保留的收缩功能和形态长达6天。还通过细胞外场电位记录证实了6天的电生理功能。这种简单的方法不仅最大限度地利用从一个单一的心脏制备的切片，从而减少了所需的动物数量，而且还通过允许多个电生理来增加数据的可重复性，药理学,生物化学，和结构研究从同一个心脏进行。
