Nguyen Doan C, Hookway Tracy A, Wu Qingling, Jha Rajneesh, Preininger Marcela K, Chen Xuemin, Easley Charles A, Spearman Paul, Deshpande Shriprasad R, Maher Kevin, Wagner Mary B, McDevitt Todd C, Xu Chunhui
Department of Pediatrics, Emory University School of Medicine and Children's Healthcare of Atlanta, Atlanta, GA 30322, USA.
Wallace H. Coulter Department of Biomedical Engineering, Georgia Institute of Technology and Emory University, Atlanta, GA 30332, USA.
Stem Cell Reports. 2014 Aug 12;3(2):260-8. doi: 10.1016/j.stemcr.2014.06.002. Epub 2014 Jul 4.
Cardiomyocytes derived from human pluripotent stem cells (hPSCs) are a promising cell source for regenerative medicine, disease modeling, and drug discovery, all of which require enriched cardiomyocytes, ideally ones with mature phenotypes. However, current methods are typically performed in 2D environments that produce immature cardiomyocytes within heterogeneous populations. Here, we generated 3D aggregates of cardiomyocytes (cardiospheres) from 2D differentiation cultures of hPSCs using microscale technology and rotary orbital suspension culture. Nearly 100% of the cardiospheres showed spontaneous contractility and synchronous intracellular calcium transients. Strikingly, from starting heterogeneous populations containing ∼10%-40% cardiomyocytes, the cell population within the generated cardiospheres featured ∼80%-100% cardiomyocytes, corresponding to an enrichment factor of up to 7-fold. Furthermore, cardiomyocytes from cardiospheres exhibited enhanced structural maturation in comparison with those from a parallel 2D culture. Thus, generation of cardiospheres represents a simple and robust method for enrichment of cardiomyocytes in microtissues that have the potential use in regenerative medicine as well as other applications.
源自人类多能干细胞（hPSC）的心肌细胞是再生医学、疾病建模和药物发现中一种很有前景的细胞来源，所有这些都需要富集的心肌细胞，理想情况下是具有成熟表型的心肌细胞。然而，目前的方法通常在二维环境中进行，这种环境会在异质群体中产生未成熟的心肌细胞。在这里，我们使用微尺度技术和旋转振荡悬浮培养从hPSC的二维分化培养物中生成了心肌细胞的三维聚集体（心肌球）。近100%的心肌球表现出自发性收缩性和同步的细胞内钙瞬变。令人惊讶的是，从起始的含有约10%-40%心肌细胞的异质群体中，生成的心肌球内的细胞群体中约80%-100%为心肌细胞，富集因子高达7倍。此外，与平行二维培养的心肌细胞相比，心肌球中的心肌细胞表现出增强的结构成熟度。因此，心肌球的生成代表了一种简单而强大的方法，可用于在微组织中富集心肌细胞，这些微组织在再生医学以及其他应用中具有潜在用途。
