Molecular analysis of the subaortic hematopoietic stem cell niche [E10.5_2]

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。该过程受到严格的时空调控，由背主动脉下方的间充质细胞等周围细胞介导。本研究结合激光显微切割与基因微阵列技术，在三个发育阶段分离背主动脉与腹主动脉组织，以探究二者的基因表达谱。具体而言，研究人员在小鼠AGM造血发生的三个阶段（E10.5、E11.5及E12.5），分别分离得到位于脊索与背主动脉顶壁之间的背侧主动脉组织，以及位于背主动脉下方的腹侧主动脉组织；经激光显微切割将组织收集至适配管盖后，使用Acturus Picopure RNA提取试剂盒完成RNA提取，并通过Affymetrix小鼠Gene 2.0 ST基因芯片开展检测。