Genetic Tagging During Human Mesoderm Differentiation Reveals Tripotent Lateral Plate Mesodermal Progenitors.
Genetic Tagging During Human Mesoderm Differentiation Reveals Tripotent Lateral Plate Mesodermal Progenitors.
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在人中胚层分化过程中的遗传标记揭示了三层侧板中胚层祖细胞。
DOI：
10.1002/stem.2351
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发表时间：
2016-05
期刊：
影响因子：
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通讯作者：
Crooks GM
中科院分区：
文献类型：
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作者：
Chin CJ;Cooper AR;Lill GR;Evseenko D;Zhu Y;He CB;Casero D;Pellegrini M;Kohn DB;Crooks GM
虽然谱系潜能的克隆研究已被广泛应用于器官特异性干细胞和祖细胞，但对早期胚胎发生中胚层形成的谱系的克隆起源知之甚少。我们应用慢病毒标记，然后通过高通量测序的载体整合位点分析（VISA）来研究造血、内皮和间充质谱系从人胚胎中胚层出现时的个体发育。与使用VISA追踪随着时间的推移显着扩增的自我更新干细胞克隆分化的研究相反，我们重点关注自我更新能力有限的祖细胞克隆群体。我们的分析揭示了采样对慢病毒标签共享解释的关键影响，特别是在具有最小克隆重复的复杂群体中。通过应用定量框架来估计采样不足的程度，我们揭示了来自多能干细胞的三能中胚层祖细胞的存在，以及它们分化成双能内皮/造血或内皮/间充质祖细胞的后续分叉。
Although clonal studies of lineage potential have been extensively applied to organ specific stem and progenitor cells, much less is known about the clonal origins of lineages formed from the germ layers in early embryogenesis. We applied lentiviral tagging followed by vector integration site analysis (VISA) with high-throughput sequencing to investigate the ontogeny of the hematopoietic, endothelial and mesenchymal lineages as they emerge from human embryonic mesoderm. In contrast to studies that have used VISA to track differentiation of self-renewing stem cell clones that amplify significantly over time, we focused on a population of progenitor clones with limited self-renewal capability. Our analyses uncovered the critical influence of sampling on the interpretation of lentiviral tag sharing, particularly among complex populations with minimal clonal duplication. By applying a quantitative framework to estimate the degree of undersampling we revealed the existence of tripotent mesodermal progenitors derived from pluripotent stem cells, and the subsequent bifurcation of their differentiation into bipotent endothelial/hematopoietic or endothelial/mesenchymal progenitors.