Department of Physiology, Xuzhou Medical University, Xuzhou, Jiangsu, China.
National Demonstration Center for Experiment Basic Medical Science Education, Xuzhou Medical University, Xuzhou, Jiangsu, China.
Biomed Mater Eng. 2022;33(5):407-421. doi: 10.3233/BME-211368.
Stem cells are used as an alternative treatment option for patients with congenital heart disease (CHD) due to their regenerative potential, but they are subject to low retention rate in the injured myocardium. Also, the diseased microenvironment in the injured myocardium may not provide healthy cues for optimal stem cell function.
In this study, we prepared a novel human-derived cardiac scaffold to improve the functional behaviors of stem cells.
Decellularized extracellular matrix (ECM) scaffolds were fabricated by removing cells of human-derived cardiac appendage tissues. Then, bone marrow c-kit+ progenitor cells from patients with congenital heart disease were seeded on the cardiac ECM scaffolds. Cell adhesion, survival, proliferation and cardiac differentiation on human cardiac decellularized ECM scaffold were evaluated in vitro. Label-free mass spectrometry was applied to analyze cardiac ECM proteins regulating cell behaviors.
It was shown that cardiac ECM scaffolds promoted stem cell adhesion and proliferation. Importantly, bone marrow c-kit+ progenitor cells cultured on cardiac ECM scaffold for 14 days differentiated into cardiomyocyte-like cells without supplement with any inducible factors, as confirmed by the increased protein level of Gata4 and upregulated gene levels of Gata4, Nkx2.5, and cTnT. Proteomic analysis showed the proteins in cardiac ECM functioned in multiple biological activities, including regulation of cell proliferation, regulation of cell differentiation, and cardiovascular system development.
The human-derived cardiac scaffold constructed in this study may help repair the damaged myocardium and hold great potential for tissue engineering application in pediatric patients with CHD.
由于干细胞具有再生潜力,因此被用作治疗先天性心脏病 (CHD) 患者的替代治疗选择,但它们在受损心肌中的保留率较低。此外,受损心肌中的病变微环境可能无法为最佳干细胞功能提供健康线索。
本研究制备了一种新型人源性心脏支架,以改善干细胞的功能行为。
通过去除人源性心脏附肢组织中的细胞来制备脱细胞细胞外基质 (ECM) 支架。然后,将来自先天性心脏病患者的骨髓 c-kit+祖细胞接种在心脏 ECM 支架上。在体外评估骨髓 c-kit+祖细胞在人心脏脱细胞 ECM 支架上的黏附、存活、增殖和心脏分化。应用无标记质谱分析调节细胞行为的心脏 ECM 蛋白。
结果表明,心脏 ECM 支架促进了干细胞的黏附和增殖。重要的是,骨髓 c-kit+祖细胞在心脏 ECM 支架上培养 14 天即可分化为心肌样细胞,而无需补充任何诱导因子,这一点通过 Gata4 蛋白水平的增加和 Gata4、Nkx2.5 和 cTnT 的基因水平上调得到证实。蛋白质组学分析表明心脏 ECM 中的蛋白质在多种生物学活性中发挥作用,包括细胞增殖的调节、细胞分化的调节和心血管系统的发育。
本研究构建的人源性心脏支架有助于修复受损的心肌,在小儿 CHD 患者的组织工程应用中具有巨大潜力。