Microarray of Mst1/2 deleted epithelial cells from E18.5 mouse lungs

ShhCre;Mst1/2flx/flx (Mst1/2 D/D) mice were generated to conditionally delete Mst1 and Mst2 from epithelial progenitors during lung morphogenesis. Lungs from E18.5 control and Mst1/2 D/D mice were mechanically and enzymatically dissociated to generate single cell suspension. Epcam(+) cells were isolated using magnetic microbeads. Microarray analysis of mRNAs isolated from Epcam(+) epithelial cells from E18.5 control and Mst1/2 D/D mice was performed to identify transcriptional changes following deletion of the mammalian Hippo kinases (Mst1 and Mst2) from the embryonic lung. The mammalian Hippo kinases, Mst1 and Mst2, were conditionally deleted in epithelial progenitors of the developing lung using ShhCre. Epcam(+) epithelial cells were isolated from the lungs of E18.5 control and Mst1/2 deleted mice. mRNA isolated from Epcam(+) epithelial cells was analyzed by microarray.本研究构建了ShhCre;Mst1/2flx/flx（Mst1/2 D/D）小鼠模型，以在肺形态发生过程中，于上皮祖细胞内条件性敲除Mst1与Mst2基因。研究人员对胚胎发育第18.5天（embryonic day 18.5，E18.5）的对照组小鼠及Mst1/2 D/D小鼠的肺组织采用机械法联合酶解法进行解离，制备得到单细胞悬液；随后通过磁微珠分选法分离得到Epcam(Epithelial Cell Adhesion Molecule, 上皮细胞黏附分子)阳性上皮细胞。通过对两组小鼠Epcam(+)上皮细胞中提取的mRNA进行基因芯片分析，本研究旨在鉴定胚胎肺上皮祖细胞中条件性敲除哺乳动物Hippo激酶Mst1与Mst2后所产生的转录组变化。 本研究借助ShhCre重组酶系统，在发育中肺的上皮祖细胞内条件性敲除Mst1与Mst2基因；从胚胎发育第18.5天的对照组小鼠及Mst1/2敲除小鼠的肺组织中分离得到Epcam(+)上皮细胞，并对其提取的mRNA开展基因芯片分析。