Notch信号通路在红系祖细胞增殖和成熟分化中的调控作用

项目名称： Notch信号通路在红系祖细胞增殖和成熟分化中的调控作用

项目编号： No.31201097

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 发育生物学与生殖生物学

项目作者： 习佳飞

作者单位： 中国人民解放军军事医学科学院

项目金额： 21万元

中文摘要： Notch信号通路参与调控造血干细胞的命运，但是notch信号通路在红系增殖和成熟分化中的作用仍不清楚。我们希望利用前期建立的造血干细胞向红系祖细胞分化及共培养诱导脱核的体系，在体外对于notch信号通路调控红系祖细胞体外增殖及成熟分化的作用进行研究，并对其作用机制进行初步的探讨。我们将研究体外不同的培养密度条件下Notch信号通路对于红系祖细胞增殖的影响，通过观察红系祖细胞表面标志的表达，分析其凋亡信号通路及增殖曲线等指标对其增殖进行分析，同时利用我们建立的共培养脱核体系分析Notch信号通路对成熟分化的影响。利用转基因的方法对红系祖细胞自身和基质细胞进行基因改造，通过这些方法确认Notch信号通路对红系祖细胞的分化中的作用。总之本研究将进一步明确Notch信号通路在红系祖细胞增殖和成熟分化中的作用，研究结果将加深我们对于红系发育分化机制的理解。

中文关键词： Notch 信号通路；红系祖细胞；细胞动力学；增殖；深度测序

英文摘要： Notch signaling regulates the fate of hematopoietic stem cells. However, the role of Notch in erythroid differentiation remains unclear. Notch signaling plays an important role in cell fate decisions in developmental systems. To clarify its role in committed hematopoietic progenitor cells, we investigated the effects of Notch signaling in erythroid progenitor cells (EPCs) generated from cord blood. In our previous studies, we established a optimized long-term serum-free culture conditions that allowed expansion into mass cultures of highly homogeneous human EPCs derived from umbilical cord blood. The mature red blood cells could be produced by co-culturing erythroid progenitors with human fetal liver stromal cells. With this procedure, it is possible to reproduce different steps of human normal erythropoiesis. The large number and purity of erythroid cells produced from a small amount of peripheral blood make this method useful for studying either normal or pathological erythropoiesis. We assayed the effects of Notch ligands on erythroid proliferation and maturation by flow cytometry, and the expression levels of the Notch target gene hairy enhancer of split (HES) -1 by real-time PCR after Notch signal activation. Overall, our study will find the pivotal role of Notch signaling in regulating erythroid maturation

英文关键词： Notch signaling pathway；erythroid progenitor cells；Cell dynamic；Proliferation；Deep sequencing