Burridge Paul W, Metzler Scott A, Nakayama Karina H, Abilez Oscar J, Simmons Chelsey S, Bruce Marc A, Matsuura Yuka, Kim Paul, Wu Joseph C, Butte Manish, Huang Ngan F, Yang Phillip C
Stanford Cardiovascular Institute Palo Alto, CA ; Department of Medicine, Division of Cardiovascular Medicine Palo Alto, CA ; Institute for Stem Cell Biology and Regenerative Medicine Palo Alto, CA.
Stanford Cardiovascular Institute Palo Alto, CA ; Department of Medicine, Division of Pediatric Cardiology Palo Alto, CA.
Am J Transl Res. 2014 Nov 22;6(6):724-35. eCollection 2014.
Therapeutic delivery of cardiomyocytes derived from human pluripotent stem cells (hPSC-CMs) represents a novel clinical approach to regenerate the injured myocardium. However, poor survival and contractility of these cells are a significant bottleneck to their clinical use. To better understand the role of cell-cell communication in enhancing the phenotype and contractile properties of hPSC-CMs, we developed a three-dimensional (3D) hydrogel composed of hPSC-CMs, human pluripotent stem cell-derived endothelial cells (hPSC-ECs), and/or human amniotic mesenchymal stem cells (hAMSCs). The objective of this study was to examine the role of multi-cellular interactions among hPSC-ECs and hAMSCs on the survival and long-term contractile phenotype of hPSC-CMs in a 3D hydrogel. Quantification of spontaneous contractility of hPSC-CMs in tri-culture demonstrated a 6-fold increase in the area of contractile motion after 6 weeks with characteristic rhythmic contraction frequency, when compared to hPSC-CMs alone (P < 0.05). This finding was supported by a statistically significant increase in cardiac troponin T protein expression in the tri-culture hydrogel construct at 6 weeks, when compared to hPSC-CMs alone (P < 0.001). The sustained hPSC-CM survival and contractility in tri-culture was associated with a significant upregulation in the gene expression of L-type Ca(2+) ion channel, Cav1.2, and the inward-rectifier potassium channel, Kir2.1 (P < 0.05), suggesting a role of ion channels in mediating these processes. These findings demonstrate that multi-cellular interactions modulate hPSC-CM phenotype, function, and survival, and they will have important implications in engineering cardiac tissues for treatment of cardiovascular diseases.
源自人多能干细胞的心肌细胞（hPSC-CMs）的治疗性递送代表了一种用于再生受损心肌的新型临床方法。然而，这些细胞的低存活率和收缩性是其临床应用的一个重大瓶颈。为了更好地理解细胞间通讯在增强hPSC-CMs的表型和收缩特性中的作用，我们开发了一种由hPSC-CMs、人多能干细胞衍生的内皮细胞（hPSC-ECs）和/或人羊膜间充质干细胞（hAMSCs）组成的三维（3D）水凝胶。本研究的目的是研究hPSC-ECs和hAMSCs之间的多细胞相互作用对3D水凝胶中hPSC-CMs的存活和长期收缩表型的作用。与单独培养的hPSC-CMs相比，三培养体系中hPSC-CMs的自发收缩性定量分析表明，6周后收缩运动面积增加了6倍，且具有特征性的节律性收缩频率（P < 0.05）。与单独培养的hPSC-CMs相比，三培养水凝胶构建体在6周时心肌肌钙蛋白T蛋白表达的统计学显著增加支持了这一发现（P < 0.001）。三培养体系中hPSC-CMs的持续存活和收缩性与L型Ca(2+)离子通道Cav1.2和内向整流钾通道Kir2.1的基因表达显著上调有关（P < 0.05），表明离子通道在介导这些过程中发挥作用。这些发现表明多细胞相互作用可调节hPSC-CMs的表型、功能和存活，并且它们将对用于治疗心血管疾病的心脏组织工程具有重要意义。
