Single cell expression profiles of the earliest cardiac precursor cells

The character of the earliest cardiac precursor cells remains largely unknown. To elucidate this further, we constructed single cell cDNAs from the mouse embryonic cardiac precursor cells of the early allantoic bud stage and the early headfold stage, and subjected them to deep sequencing. The most anterior part of the embryos where cardiac precursor cells exist was digested by trypsin to separate into single cells. After a cell was transferred into a reaction tube, single cell cDNAs were constructed as PCR amplicons. cDNAs of cardiac precursor cells were identified by PCR of marker genes.最早的心脏前体细胞（cardiac precursor cells）的特征在很大程度上仍未明确。为进一步阐明该科学问题，我们从处于早期尿囊芽阶段（early allantoic bud stage）及早期头褶阶段（early headfold stage）的小鼠胚胎心脏前体细胞中构建了单细胞cDNA，并对其开展深度测序（deep sequencing）。实验过程中，我们首先将存在心脏前体细胞的胚胎最前部区域经胰蛋白酶消化解离为单个细胞；随后将单个细胞转移至反应管，以PCR扩增子（PCR amplicons）的形式构建单细胞cDNA；最后通过标记基因的聚合酶链式反应（PCR）扩增，鉴定得到心脏前体细胞的cDNA。