Department of Cell and Tissue Engineering, Fraunhofer Institute for Interfacial Engineering and Biotechnology IGB, Stuttgart 70569, Germany; Department of Women's Health, Research Institute of Women's Health, University Hospital of the Eberhard Karls University Tübingen, Tübingen 72076, Germany.
Department of Cell and Tissue Engineering, Fraunhofer Institute for Interfacial Engineering and Biotechnology IGB, Stuttgart 70569, Germany.
Stem Cell Reports. 2017 Jul 11;9(1):122-135. doi: 10.1016/j.stemcr.2017.04.021. Epub 2017 May 18.
Cardiovascular disease remains a leading cause of mortality and morbidity worldwide. Embryonic stem cell-derived cardiomyocytes (ESC-CMs) may offer significant advances in creating in vitro cardiac tissues for disease modeling, drug testing, and elucidating developmental processes; however, the induction of ESCs to a more adult-like CM phenotype remains challenging. In this study, we developed a bioreactor system to employ pulsatile flow (1.48 mL/min), cyclic strain (5%), and extended culture time to improve the maturation of murine and human ESC-CMs. Dynamically-cultured ESC-CMs showed an increased expression of cardiac-associated proteins and genes, cardiac ion channel genes, as well as increased SERCA activity and a Raman fingerprint with the presence of maturation-associated peaks similar to primary CMs. We present a bioreactor platform that can serve as a foundation for the development of human-based cardiac in vitro models to verify drug candidates, and facilitates the study of cardiovascular development and disease.
心血管疾病仍然是全球范围内导致死亡和发病的主要原因。胚胎干细胞衍生的心肌细胞 (ESC-CMs) 可能在创建用于疾病建模、药物测试和阐明发育过程的体外心脏组织方面提供重大进展;然而,将 ESC 诱导为更类似于成人的 CM 表型仍然具有挑战性。在这项研究中,我们开发了一种生物反应器系统,采用脉动流(1.48 毫升/分钟)、循环应变(5%)和延长培养时间来改善鼠和人 ESC-CMs 的成熟度。动态培养的 ESC-CMs 表现出心脏相关蛋白和基因、心脏离子通道基因的表达增加,以及 SERCA 活性增加和拉曼指纹的出现,其中存在与成熟相关的峰类似于原代心肌细胞。我们提出了一个生物反应器平台,可作为开发基于人类的体外心脏模型的基础,以验证候选药物,并促进心血管发育和疾病的研究。