Vink Chris Sebastiaan, Calero-Nieto Fernando Jose, Wang Xiaonan, Maglitto Antonio, Mariani Samanta Antonella, Jawaid Wajid, Göttgens Berthold, Dzierzak Elaine
Centre for Inflammation Research, Queens Medical Research Institute, University of Edinburgh, Edinburgh, Midlothian, Scotland EH16 4TJ, UK.
Department of Haematology, Wellcome & MRC Cambridge Stem Cell Institute, University of Cambridge, Cambridge, Cambridgeshire, England CB2 0AW, UK.
Cell Rep. 2020 May 12;31(6):107627. doi: 10.1016/j.celrep.2020.107627.
Whereas hundreds of cells in the mouse embryonic aorta transdifferentiate to hematopoietic cells, only very few establish hematopoietic stem cell (HSC) identity at a single time point. The Gata2 transcription factor is essential for HSC generation and function. In contrast to surface-marker-based cell isolation, Gata2-based enrichment provides a direct link to the internal HSC regulatory network. Here, we use iterations of index-sorting of Gata2-expressing intra-aortic hematopoietic cluster (IAHC) cells, single-cell transcriptomics, and functional analyses to connect HSC identity to specific gene expression. Gata2-expressing IAHC cells separate into 5 major transcriptomic clusters. Iterative analyses reveal refined CD31, cKit, and CD27 phenotypic parameters that associate specific molecular profiles in one cluster with distinct HSC and multipotent progenitor function. Thus, by iterations of single-cell approaches, we identify the transcriptome of the first functional HSCs as they emerge in the mouse embryo and localize them to aortic clusters containing 1-2 cells.
在小鼠胚胎主动脉中，数百个细胞可转分化为造血细胞，但在单个时间点，只有极少数细胞能建立造血干细胞（HSC）身份。Gata2转录因子对造血干细胞的产生和功能至关重要。与基于表面标志物的细胞分离方法不同，基于Gata2的富集方法直接连接到内部造血干细胞调控网络。在这里，我们通过对表达Gata2的主动脉内造血簇（IAHC）细胞进行索引排序、单细胞转录组学和功能分析的迭代，将造血干细胞身份与特定基因表达联系起来。表达Gata2的IAHC细胞可分为5个主要的转录组簇。迭代分析揭示了精细的CD31、cKit和CD27表型参数，这些参数将一个簇中的特定分子谱与不同的造血干细胞和多能祖细胞功能联系起来。因此，通过单细胞方法的迭代，我们确定了小鼠胚胎中最早出现的功能性造血干细胞的转录组，并将它们定位到含有1-2个细胞的主动脉簇中。
