Co-Differentiation of Human Pluripotent Stem Cells-Derived Cardiomyocytes and Endothelial Cells from Cardiac Mesoderm Provides a Three-Dimensional Model of Cardiac Microtissue.
Co-Differentiation of Human Pluripotent Stem Cells-Derived Cardiomyocytes and Endothelial Cells from Cardiac Mesoderm Provides a Three-Dimensional Model of Cardiac Microtissue.
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DOI：
10.1002/cphg.46
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发表时间：
2017-10-18
影响因子：
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通讯作者：
Mummery, Christine L
中科院分区：
文献类型：
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作者：
Giacomelli, Elisa;Bellin, Milena;Mummery, Christine L
心脏发育过程中，心脏中胚层亚型的形成在时间和空间上受到高度调节。基于人多能干细胞（hPS细胞）的体外模型提供了研究人类胚胎中常见心血管祖细胞谱系特化的命运选择机制的机会。特别是心脏内皮细胞的产生允许创建复杂的心血管疾病模型，其中心肌细胞或内皮细胞受到影响。在这里，描述了使用hPS细胞从心脏中胚层共分化心肌细胞和内皮细胞的方案。从异质分化培养物中富集每种细胞群的精确细节，如何维持和分离富集的心肌细胞的描述，以及富集的内皮细胞的扩增和冷冻保存都被提供。从这些细胞群的三维心脏微组织的生成和培养进行了描述，并提供了通过免疫荧光染色和重新铺板用于下游应用的微组织表征指南。John Wiley & Sons，Inc.
The formation of cardiac mesodermal subtypes is highly regulated in time and space during heart development. In vitro models based on human pluripotent stem cells (hPS cells) provide opportunities to study mechanisms underlying fate choices governing lineage specification from common cardiovascular progenitors in human embryos. The generation of cardiac endothelial cells in particular allows the creation of complex models of cardiovascular disorders in which either cardiomyocytes or endothelial cells are affected. Here, a protocol for co-differentiation of cardiomyocytes and endothelial cells from cardiac mesoderm using hPS cells is described. Precise details for the enrichment of each cell population from heterogeneous-differentiated cultures, a description of how to maintain and dissociate enriched cardiomyocytes, and the expansion and cryopreservation of enriched endothelial cells are all provided. The generation and culture of three-dimensional cardiac microtissues from these cell populations is described and guidelines for the characterization of microtissues by immunofluorescent staining and re-plating for downstream applications are provided. © 2017 by John Wiley & Sons, Inc.