Organotypic slice culture from human adult ventricular myocardium
Organotypic slice culture from human adult ventricular myocardium
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DOI：
10.1093/cvr/cvr259
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发表时间：
2012-01-01
影响因子：
10.8
通讯作者：
Dendorfer, Andreas
中科院分区：
文献类型：
--
作者：
Brandenburger, Matthias;Wenzel, Jan;Dendorfer, Andreas
心血管研究需要复杂的和功能完整的实验模型。由于物种之间心肌的细胞和亚细胞组成存在重大差异，因此非常需要使用人类心脏组织。为了提高实验使用的人类心肌，我们建立了方法，从成人心室肌的重要组织切片的制备以及他们的长期保存在器官型culture.Methods和结果的条件人心室的心脏样本来自手术标本切除治疗Morrow myectomy和切成300毫米厚的切片。切片在急性实验中表征或在液-气界面处培养。通过活力染色、酶活性试验、细胞内电位记录和力测量证明活力和功能性。精密切割的切片在培养的28天内表现出高活力，并显示出典型的心肌细胞动作电位特征，这使得在整个培养期间对延迟整流钾电流（I（Kr））和ATP依赖性钾通道的快速组分进行药理学安全性测试成为可能。通过定量PCR证实主要离子通道的恒定表达。急性切片产生兴奋依赖性收缩，具有明显的前负荷依赖性和β-肾上腺素能反应。收缩力和肌球蛋白轻链的表达下降，在培养的第一天，但达到稳定状态与反应后，β-肾上腺素能刺激被preserved.Conclusion器官型心脏切片代表了一个多细胞模型的人类心肌和一个新的平台，从分子相互作用的研究，组织工程。
Aims Cardiovascular research requires complex and functionally intact experimental models. Due to major differences in the cellular and subcellular composition of the myocardium between species, the use of human heart tissue is highly desirable. To enhance the experimental use of the human myocardium, we established methods for the preparation of vital tissue slices from the adult ventricular myocardium as well as conditions for their long-term preservation in organotypic culture.Methods and results Human ventricular heart samples were derived from surgical specimens excised during a therapeutic Morrow myectomy and cut into 300 mm thick slices. Slices were either characterized in acute experiments or cultured at a liquid-air interface. Viability and functionality were proven by viability staining, enzyme activity tests, intracellular potential recordings, and force measurements. Precision-cut slices showed high viability throughout 28 days in culture and displayed typical cardiomyocyte action potential characteristics, which enabled pharmacological safety testing on the rapid component of the delayed rectifier potassium current (I(Kr)) and ATP-dependent potassium channels throughout the whole culture period. Constant expression of major ion channels was confirmed by quantitative PCR. Acute slices developed excitation-dependent contractions with a clear preload dependency and a beta-adrenergic response. Contractility and myosin light chain expression decreased during the first days in culture but reached a steady state with reactivity upon beta-adrenergic stimulation being preserved.Conclusion Organotypic heart slices represent a multicellular model of the human myocardium and a novel platform for studies ranging from the investigation of molecular interactions to tissue engineering.