论文部分内容阅读
目的 :Notch信号通路对造血干 /祖细胞的增殖分化具有重要调控作用。Delta like 1(Dll 1)是Notch的配体之一 ,本研究观察了重组人Dll 1胞外区 (hDll 1ext)对细胞因子扩增脐带血造血祖细胞的促进作用。方法 :RT PCR用于检测Notch 1及其配体在脐带血细胞上的表达情况。利用免疫磁性分离 (MACS)的方法分离人脐带血CD34+ 细胞。在无血清培养体系中 ,hDll 1ext分别和不同的细胞因子组合联合培养CD34+ 细胞 ,通过集落形成实验检测hDll 1ext对造血祖细胞的扩增作用。结果 :RT PCR方法检测到脐带血单个核细胞表达Notch 1和它的配体Dll 1,Jagged 1,表明在生理条件下 ,脐血细胞的增殖分化可能受到Notch信号的调控。hDll 1ext能促进细胞因子组合对脐带血HPP CFC的扩增 ,其中hDll 1ext和SCF ,IL 3,IL 6联合的扩增作用最强 ,无血清培养 8,12d后 ,HPP CFC分别增加为培养前的 9.7和 4 3倍。结论 :重组hDll 1ext能促进细胞因子对脐带血原始造血祖细胞的扩增作用
Objective: Notch signaling pathway plays an important regulatory role in the proliferation and differentiation of hematopoietic stem / progenitor cells. Delta like 1 (Dll 1) is one of the ligands of Notch. In this study, we investigated the effects of hD11 lectin on the proliferation of umbilical cord blood-derived hematopoietic progenitor cells by cytokines. Methods: RT PCR was used to detect the expression of Notch 1 and its ligand on umbilical cord blood cells. Human umbilical cord blood CD34 + cells were isolated by immunomagnetic separation (MACS). In serum-free culture system, hD11ext and CD34 + cells were cultured with different combinations of cytokines respectively. The colony formation assay was used to detect the amplification of hematopoietic progenitor cells by hD11ext. Results: Notch 1 and its ligands Dll 1 and Jagged 1 were detected by RT-PCR in cord blood mononuclear cells, indicating that under physiological conditions, the proliferation and differentiation of cord blood cells may be regulated by Notch signaling. hDll 1ext could promote the expansion of HPC CFCs in umbilical cord blood by cytokine combination. The combination of hDll 1ext and SCF, IL 3 and IL 6 had the strongest amplification effect. After 8 and 12 days of serum-free culture, The 9.7 and 4 3 times. CONCLUSION: Recombinant hD11ext can promote the expansion of cord blood progenitor cells by cytokines