Llucià-Valldeperas Aida, Bragós Ramon, Bayés-Genís Antoni
Insuficiencia Cardiaca y Regeneración Cardiaca (ICREC) Research Program, Health Science Research Institute Germans Trias i Pujol; Amsterdam Universitair Medisch Centrum (UMC), Vrije Universiteit Amsterdam, Pulmonology and Physiology, Amsterdam Cardiovascular Sciences;
Electronic and Biomedical Instrumentation Group, Departament d'Enginyeria Electrònica, Universitat Politècnica de Catalunya.
J Vis Exp. 2019 Jan 18(143). doi: 10.3791/58934.
Cardiovascular diseases are the leading cause of death in developed countries. Consequently, the demand for effective cardiac cell therapies has motivated researchers in the stem cell and bioengineering fields to develop in vitro high-fidelity human myocardium for both basic research and clinical applications. However, the immature phenotype of cardiac cells is a limitation on obtaining tissues that functionally mimic the adult myocardium, which is mainly characterized by mechanical and electrical signals. Thus, the purpose of this protocol is to prepare and mature the target cell population through electromechanical stimulation, recapitulating physiological parameters. Cardiac tissue engineering is evolving toward more biological approaches, and strategies based on biophysical stimuli, thus, are gaining momentum. The device developed for this purpose is unique and allows individual or simultaneous electrical and mechanical stimulation, carefully characterized and validated. In addition, although the methodology has been optimized for this stimulator and a specific cell population, it can easily be adapted to other devices and cell lines. The results here offer evidence of the increased cardiac commitment of the cell population after electromechanical stimulation. Electromechanically stimulated cells show an increased expression of main cardiac markers, including early, structural, and calcium-regulating genes. This cell conditioning could be useful for further regenerative cell therapy, disease modeling, and high-throughput drug screening.
心血管疾病是发达国家的主要死因。因此，对有效的心脏细胞疗法的需求促使干细胞和生物工程领域的研究人员开发用于基础研究和临床应用的体外高保真人类心肌。然而，心脏细胞的未成熟表型是获得功能上模拟成年心肌组织的一个限制因素，成年心肌主要以机械和电信号为特征。因此，本方案的目的是通过机电刺激使目标细胞群体成熟，概括生理参数。心脏组织工程正朝着更具生物学特性的方法发展，因此基于生物物理刺激的策略正获得发展动力。为此目的开发的装置是独特的，允许单独或同时进行电刺激和机械刺激，并经过仔细表征和验证。此外，尽管该方法已针对此刺激器和特定细胞群体进行了优化，但它可以很容易地适用于其他装置和细胞系。此处的结果提供了证据，证明机电刺激后细胞群体的心脏定向分化增加。经机电刺激的细胞显示出主要心脏标志物的表达增加，包括早期、结构和钙调节基因。这种细胞预处理可能对进一步的再生细胞治疗、疾病建模和高通量药物筛选有用。
