本文選題：內(nèi)皮細胞 + 細胞培養(yǎng)技術　； 參考：《華中科技大學》2008年碩士論文

【摘要】： 背景外科手術中牽拉鉗夾所致的血管內(nèi)膜損傷,將加重缺血再灌注損傷,加速血栓形成,是直接影響胸心外科術后病變組織的血流灌注情況的關鍵。研究證實內(nèi)皮祖細胞(endothelial progenitor cells,EPCs)可以誘導、調節(jié)組織缺血缺氧區(qū)血管新生和血管生成,刺激損傷血管內(nèi)皮重新愈合,從而改善缺血器官功能。 目的本實驗根據(jù)目前比較公認的EPCs表面特異性分子標志(CD34~+/CD133~+/vWF~+)從臍血中分離出EPCs,并在體外進行誘導分化,利用臍血在體外血管器官培養(yǎng)基礎上觀察了EPCs移植對去內(nèi)膜血管段的修復過程。 方法采用磁珠分選法及培養(yǎng)人臍血內(nèi)皮祖細胞,分別采用流式細胞術及因子相關抗原免疫染色對培養(yǎng)細胞進行鑒定。牽拉鉗夾損傷法制備去內(nèi)膜臍動脈段,與EPCs共孵育7d后取內(nèi)皮祖細胞移植組和對照組血管段,采用病理學方法、免疫組織化學和圖像分析技術評價內(nèi)皮祖細胞的修復效果。 結果體外成功培養(yǎng)出人臍血內(nèi)皮祖細胞,流式細胞術分析顯示培養(yǎng)7d后,Ⅷ因子相關抗原(vWF)、白細胞分化抗原34(CD34)免疫染色陽性,流式細胞術顯示我們的細胞表達EPCs最特異性早期抗原AC133 90%以上。人臍血EPCs與去內(nèi)膜臍動脈共培育使之再內(nèi)皮化,形成新內(nèi)膜。 結論利用臍血可成功培養(yǎng)出內(nèi)皮祖細胞,體外培養(yǎng)內(nèi)皮祖細胞可修復內(nèi)皮損傷血管。
[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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