Expression data from developing mouse livers

Roles of mesothelial cells (MCs) are poorly understood during liver development and injury. We identified podoplanin (Pdpn) as a cell surface markers for mesothelial cells in E12.5 mouse developing liver. To identify genes uniquely expressed in MCs, we isolated MCs from E12.5 mouse livers by FACS using anti-Pdpn antibodies and performed microarray analysis. The E12.5 liver was digested with trypsin-EDTA and incubated with antibodies against Pdpn. MCs were isolated as Pdpn+ population by FACS. Total RNA was extracted with RNAqueous Micro (Ambion) and the probes for the microarray were synthesized using the Ovation RNA amplification system V2 and FL-Ovation cDNA Biotin module V2 (Nugen). The labeled probes were hybridized with GeneChip Mouse Genome 430 2.0 arrays (Affymetrix) and signals were analyzed with Genomic Suite software (Partek).间皮细胞（mesothelial cells, MCs）在肝脏发育与损伤过程中的生物学功能迄今尚未明确。本研究鉴定出足突蛋白（podoplanin, Pdpn）可作为E12.5小鼠发育肝脏中间皮细胞的细胞表面标记物。为筛选仅在间皮细胞中特异性表达的基因，我们通过荧光激活细胞分选术（Fluorescence-Activated Cell Sorting, FACS），利用抗Pdpn抗体从E12.5小鼠肝脏中分离得到间皮细胞，并开展微阵列分析。实验人员先用胰蛋白酶-EDTA消化E12.5小鼠肝脏组织，随后将其与抗Pdpn抗体孵育，通过荧光激活细胞分选术分离获得Pdpn阳性的间皮细胞群。使用RNAqueous Micro试剂盒（Ambion公司）提取总RNA，并借助Ovation RNA扩增系统V2与FL-Ovation cDNA生物素标记模块V2（Nugen公司）合成微阵列探针。将标记完成的探针与GeneChip小鼠基因组430 2.0芯片（Affymetrix公司）进行杂交，随后采用Genomic Suite软件（Partek公司）对杂交信号进行分析。