iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901 Oeiras, Portugal.
Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal.
ACS Appl Bio Mater. 2021 Feb 15;4(2):1888-1899. doi: 10.1021/acsabm.0c01490. Epub 2021 Jan 29.
Human induced pluripotent stem cells (hiPSC) possess significant therapeutic potential due to their high self-renewal capability and potential to differentiate into specialized cells such as cardiomyocytes. However, generated hiPSC-derived cardiomyocytes (hiPSC-CM) are still immature, with phenotypic and functional features resembling the fetal rather than their adult counterparts, which limits their application in cell-based therapies, cardiac disease modeling, and drug cardiotoxicity screening. Recent discoveries have demonstrated the potential of the extracellular matrix (ECM) as a critical regulator in development, homeostasis, and injury of the cardiac microenvironment. Within this context, this work aimed to assess the impact of human cardiac ECM in the phenotype and maturation features of hiPSC-CM. Human ECM was isolated from myocardium tissue through a physical decellularization approach. The cardiac tissue decellularization process reduced DNA content significantly while maintaining ECM composition in terms of sulfated glycosaminoglycans (s-GAG) and collagen content. These ECM particles were successfully incorporated in three-dimensional (3D) hiPSC-CM aggregates (CM+ECM) with no impact on viability and metabolic activity throughout 20 days in 3D culture conditions. Also, CM+ECM aggregates displayed organized and longer sarcomeres, with improved calcium handling when compared to hiPSC-CM aggregates. This study shows that human cardiac ECM functionalization of hiPSC-based cardiac tissues improves cardiomyocyte maturation. The knowledge generated herein provides essential insights to streamline the application of ECM in the development of hiPSC-based therapies targeting cardiac diseases.
人诱导多能干细胞（hiPSC）具有高自我更新能力和分化为特定细胞（如心肌细胞）的潜力，因此具有重要的治疗潜力。然而，生成的 hiPSC 衍生心肌细胞（hiPSC-CM）仍然不成熟，其表型和功能特征类似于胎儿，而不是其成年对应物，这限制了它们在基于细胞的治疗、心脏疾病建模和药物心脏毒性筛选中的应用。最近的发现表明细胞外基质（ECM）作为心脏微环境发育、稳态和损伤的关键调节剂具有潜力。在这种情况下，本工作旨在评估人心脏 ECM 对 hiPSC-CM 表型和成熟特征的影响。人心脏 ECM 通过物理去细胞化方法从心肌组织中分离出来。心脏组织的去细胞化过程显著降低了 DNA 含量，同时保持了 ECM 成分中硫酸化糖胺聚糖（s-GAG）和胶原蛋白含量。这些 ECM 颗粒成功地整合到三维（3D）hiPSC-CM 聚集体（CM+ECM）中，在 3D 培养条件下 20 天内对活力和代谢活性没有影响。此外，与 hiPSC-CM 聚集体相比，CM+ECM 聚集体显示出更有序和更长的肌节，以及改善的钙处理。本研究表明，人心脏 ECM 对基于 hiPSC 的心脏组织的功能化可改善心肌细胞成熟。本文所产生的知识为简化 ECM 在针对心脏疾病的基于 hiPSC 的治疗开发中的应用提供了重要的见解。
