A spatiotemporal cell atlas of cardiopulmonary progenitor cell allocation during development

The mesoderm surrounding both the developing heart and anterior foregut endoderm provides instructive cues guiding lung development and serves as a source of cardiopulmonary mesodermal progenitor cells (CPPs) expressing Wnt2 that give rise to both cardiac inflow tract and lung mesodermal cell lineages. To assess mechanisms of Wnt2+ CPP contribution to heart and lung development, we performed single cell RNA sequencing (scRNA-seq) on lineage-traced cells labeled at E8.5 and collected their progeny across the developmental lifespan. Wnt2-CreERT2 was used to drive lineage expression of EYFP. Lung and heart cells positive for EYFP were isolated by FACS at multiple developmental timepoints.

围绕发育中心脏及前肠前部内胚层的中胚层，可提供指导肺发育的诱导性信号，同时亦是表达Wnt2的心肺中胚层祖细胞（cardiopulmonary mesodermal progenitor cells, CPPs）的来源——这类祖细胞可分化为心脏流入道与肺的中胚层细胞谱系。为探究表达Wnt2的CPPs对心脏与肺发育的贡献机制，我们对胚胎第8.5天（E8.5）标记的谱系示踪细胞开展了单细胞RNA测序（single cell RNA sequencing, scRNA-seq），并在整个发育周期中收集其子代细胞。我们使用Wnt2-CreERT2驱动增强型黄色荧光蛋白（enhanced yellow fluorescent protein, EYFP）的谱系表达，并在多个发育时间点通过荧光激活细胞分选术（Fluorescence-Activated Cell Sorting, FACS）分离得到EYFP阳性的肺与心脏细胞。