Guiding T lymphopoiesis from pluripotent stem cells by defined transcription factors [single hematopoietic cells (iHEC)]

We identified that the coordinated expression of exogenous Runx1 and Hoxa9 during the haematopoietic induction process from mouse PSC resulted in a type of induced hemogenic endothelial cells (iHEC). The single iHEC can be robustly educated into induced haematopoietic progenitor cells (iHPC), which gave rise to induced T cells (iT cells) with abundant TCRαβ repertoire in vivo. We sorted the single iHEC and iHPC from day-14, day-17, day-21 cell products derived from 241 iR9-ES and performed single-cell RNA-Seq. mRNA profiles of 73 single hematopoietic endothelia cells (iHEC). single HCs were sorted from PSC-derived cells for RNAseq analysis according to kit instruction of Discover-sc WTA Kit V2(Vazyme) and TruePrepTM  DNA Library Prep Kit V2 (Vazyme), using illumina NextSeq500.

本研究证实，在小鼠多能干细胞（PSC）的造血诱导过程中，外源性Runx1与Hoxa9的协同表达可诱导产生一类诱导性造血内皮细胞（induced hemogenic endothelial cells，iHEC）。单个诱导性造血内皮细胞可高效定向诱导分化为诱导性造血祖细胞（induced haematopoietic progenitor cells，iHPC），后者在体内可分化产生具有丰富T细胞受体αβ（TCRαβ）库的诱导性T细胞（induced T cells，iT细胞）。本研究从241株iR9-ES细胞诱导得到的第14天、第17天、第21天细胞产物中分选得到单个iHEC与iHPC，并开展了单细胞RNA测序，共获得73个单个诱导性造血内皮细胞的mRNA表达谱。此外，研究人员依照Vazyme公司的Discover-sc WTA Kit V2与TruePrep™ DNA Library Prep Kit V2试剂盒说明书，从多能干细胞诱导得到的细胞中分选得到单个造血细胞，采用Illumina NextSeq500测序平台完成RNA测序分析。