Molecular analysis of the subaortic hematopoietic stem cell niche [E11.5]

Hematopoietic stem cells (HSCs) are formed in the Aorta-Gonad-Mesobephros (AGM) region of mouse midgestation embryos. This process is tightly regulated in time and space by surrounding cells inculding mesenchymal cells located underneath the dorsal aorta. We combined laser microdissection and microarrays to isolate the dorsal and ventral aortic tissues at three developmental stages and to explore their gene expression profiling. The dorsal and ventral aortic tissues (located in between the notochord and the roof of the dorsal aorta and underneath the dorsal aorta, respectively) were isolated by laser microdissection in the mouse embryo at three stages of AGM hematopoiesis (E10.5, E11.5 and E12.5). Microdissected tissues were collected on appropriate caps and then proceeded for RNA extraction using Acturus Picopure RNA extraction kit and Affymetrix microarrays mouse Gene 2.0 ST.造血干细胞（Hematopoietic stem cells, HSCs）形成于小鼠妊娠中期胚胎的主动脉-性腺-中肾（Aorta-Gonad-Mesobephros, AGM）区域。该过程受包括背主动脉下方间充质细胞在内的周围细胞在时间与空间维度上的严格调控。本研究结合激光显微切割与基因芯片技术，在三个发育阶段分离背侧与腹侧主动脉组织，以探究其基因表达谱。在小鼠AGM造血发生的三个阶段（E10.5、E11.5及E12.5）的胚胎中，研究人员通过激光显微切割分别分离出位于脊索与背主动脉顶壁之间，以及背主动脉下方的背侧主动脉组织与腹侧主动脉组织。将显微切割获取的组织收集至专用管盖后，采用Acturus Picopure RNA提取试剂盒完成RNA提取，并使用Affymetrix小鼠Gene 2.0 ST基因芯片开展后续检测。