人脐血EPCs修复脐动脉内膜损伤的实验研究

发布时间：2018-06-27 12:16

  本文选题：内皮细胞 + 细胞培养技术　； 参考：《华中科技大学》2008年硕士论文

【摘要】： 背景外科手术中牵拉钳夹所致的血管内膜损伤,将加重缺血再灌注损伤,加速血栓形成,是直接影响胸心外科术后病变组织的血流灌注情况的关键。研究证实内皮祖细胞(endothelial progenitor cells,EPCs)可以诱导、调节组织缺血缺氧区血管新生和血管生成,刺激损伤血管内皮重新愈合,从而改善缺血器官功能。 目的本实验根据目前比较公认的EPCs表面特异性分子标志(CD34~+/CD133~+/vWF~+)从脐血中分离出EPCs,并在体外进行诱导分化,利用脐血在体外血管器官培养基础上观察了EPCs移植对去内膜血管段的修复过程。 方法采用磁珠分选法及培养人脐血内皮祖细胞,分别采用流式细胞术及因子相关抗原免疫染色对培养细胞进行鉴定。牵拉钳夹损伤法制备去内膜脐动脉段,与EPCs共孵育7d后取内皮祖细胞移植组和对照组血管段,采用病理学方法、免疫组织化学和图像分析技术评价内皮祖细胞的修复效果。 结果体外成功培养出人脐血内皮祖细胞,流式细胞术分析显示培养7d后,Ⅷ因子相关抗原(vWF)、白细胞分化抗原34(CD34)免疫染色阳性,流式细胞术显示我们的细胞表达EPCs最特异性早期抗原AC133 90%以上。人脐血EPCs与去内膜脐动脉共培育使之再内皮化,形成新内膜。 结论利用脐血可成功培养出内皮祖细胞,体外培养内皮祖细胞可修复内皮损伤血管。
[Abstract]:Background the injury of vascular intima caused by pulling clamp in surgical operation will aggravate the injury of ischemia reperfusion and accelerate the formation of thrombus. It is the key to directly affect the blood flow perfusion of the pathological tissue after thoracic and cardiac surgery. It is demonstrated that endothelial progenitor cells (endothelial progenitor cells) can induce angiogenesis and angiogenesis in ischemic and hypoxic areas and stimulate endothelial rehealing to improve the function of ischemic organs. Objective to isolate EPCs from cord blood according to the known surface specific molecular markers of EPCs (CD34 ~ / CD133 ~ / vWF~), and to induce EPCs to differentiate in vitro. On the basis of vascular organ culture in vitro, umbilical cord blood was used to observe the repair process of endarterectomy vascular segment by EPCs transplantation. Methods Human umbilical cord blood endothelial progenitor cells were isolated by magnetic beads and identified by flow cytometry and factor associated antigen immunostaining. After incubating with EPCs for 7 days, endothelial progenitor cells (EPCs) were harvested from vascular segments of endothelial progenitor cells transplantation group and control group. The repair effect of endothelial progenitor cells was evaluated by histopathology, immunohistochemistry and image analysis. Results Human umbilical cord blood endothelial progenitor cells were successfully cultured in vitro. Flow cytometry showed that the factor 鈪，

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