A Rainbow Reporter Tracks Single Cells and Reveals Heterogeneous Cellular Dynamics among Pluripotent Stem Cells and Their Differentiated Derivatives
A Rainbow Reporter Tracks Single Cells and Reveals Heterogeneous Cellular Dynamics among Pluripotent Stem Cells and Their Differentiated Derivatives
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DOI：
10.1016/j.stemcr.2020.06.005
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发表时间：
2020-07-14
影响因子：
5.9
通讯作者：
Davis, Jennifer
中科院分区：
文献类型：
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作者：
El-Nachef, Danny;Shi, Kevin;Davis, Jennifer
单细胞转录组学方法已经在多能干细胞（PSCs）群体中发现了分子异质性。一种追踪单细胞谱系及其纵向表型的工具将揭示异质性是否超出分子身份。因此，我们产生了一个稳定的可诱导的彩虹报告人PSC系，提供多达18个独特的膜靶向荧光条形码。这些条形码可以重复评估单个细胞，因为它们克隆扩展，改变形态和迁移。由于该报告的细胞分辨率，我们鉴定出具有增强克隆扩增，同步细胞分裂和持续定位到集合体边缘的PSCs亚群。在定向分化为心肌细胞、皮质神经元和肝母细胞的过程中，报告基因的表达稳定维持。神经分化的反复检查显示，自组装的皮质组织来源于克隆优势祖细胞。总的来说，这些发现证明了该报告线用于跟踪单细胞行为的广泛实用性和易于实现。
Single-cell transcriptomic approaches have found molecular heterogeneities within populations of pluripotent stem cells (PSCs). A tool that tracks single-cell lineages and their phenotypes longitudinally would reveal whether heterogeneity extends beyond molecular identity. Hence, we generated a stable Cre-inducible rainbow reporter human PSC line that provides up to 18 unique membrane-targeted fluorescent barcodes. These barcodes enable repeated assessments of single cells as they clonally expand, change morphology, and migrate. Owing to the cellular resolution of this reporter, we identified subsets of PSCs with enhanced clonal expansion, synchronized cell divisions, and persistent localization to colony edges. Reporter expression was stably maintained throughout directed differentiation into cardiac myocytes, cortical neurons, and hepatoblasts. Repeated examination of neural differentiation revealed self-assembled cortical tissues derive from clonally dominant progenitors. Collectively, these findings demonstrate the broad utility and easy implementation of this reporter line for tracking single-cell behavior.