人胚胎造血的AGM区基质细胞基因表达谱分析VIP专享

人胚胎造血的 AGM 区基质细胞基因表达谱分析【摘要】 为了筛选在主动脉-性腺-中肾区基质细胞中差异表达的基因，以药物流产的孕 30-35 天的人胚胎 AGM 区来源基质细胞为“实验方”，以源自意外而致流产的孕 4-5 月的水囊引产的胎儿肝组织的基质细胞为“驱动方”，建立了在人 AGM区基质细胞中差异表达基因的消减杂交文库。进一步用基因芯片技术对所建立的抑制消减杂交文库进行筛选，并选择其中明显差异表达的 18 个克隆进行序列测定和同源性分析。结果表明： 在所构建的 AGM 抑制消减杂交文库中，经过 PCR 鉴定有特异性扩增带且扩增带分子量在 200-700 bp 的克隆有 211 个，阳性率高达％。经过基因芯片筛选，选择出 ratio 值大于 5 的克隆共18 个进行序列测定显示，在测序的 18 个明显差异表达的阳性克隆中，4 个为未知基因，14 个为已知基因，其中这些已知基因分别参加了细胞迁移、分化、生长、细胞周期、信号转导及血管发生等细胞重要生命过程。这些基因大多数在胚胎分化造血发育中的作用尚未见报道。结论 筛选 AGM 区基质细胞中差异表达的基因为进一步讨论胚胎造血发育的分化调控的分子机理提供了必要的理论基础。【关键词】 AGM 区；胎肝；基质细胞；抑制消减杂交；基因芯片 Screening and Cloning the Genes Differentially Expressed in Human Embryonic AGM-Derived Stromal Cells Abstract To screen and separate the genes differentially expressed in human embryonic aorta-gonad-mesonephros(AGM)-derived stromal cells，a subtracted library was generated through the suppression subtractive hybridization using the cDNA of human embryonic AGM-derived stromal cells as target and human fetal liver(FL)-derived stromal cells as drivers. Then a high though screening technique，gene chip，was used to screen the differentially expressed genes in the established subtractive library. Approximately 18 of the resulting subtracted cDNA clones were partially sequenced and analyzed by blastn in the GenBank database. The results showed that 211 Clones were selected and identified from the established subtractive library，the positive ...