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自体骨髓CD34~+干细胞移植治疗严重肢体缺血的实验研究

Therapeutic neovascularization with autologous bone marrow CD34~+ cells transplantation in hindlimb ischemia

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【作者】 张鸿坤张楠吴丽花金炜李鸣封华赵海格陈旭东杨春虎

【Author】 ZHANG Hong-kun*, ZHANG Nan, WU Li-hua, JIN Wei, LI Ming, FENG Hua, ZHAO Hai-ge, CHEN Xu-dong, YANG Chun-hu. *Department of Vascular Surgery, the First Affiliated Hospital,College of Medicine, Zhejiang University, Hangzhou 310003, China

【机构】 浙江大学医学院附属第一医院血管外科浙江大学医学院附属邵逸夫医院内分泌科浙江大学医学院生物电磁学省重点实验室 310003杭州310003杭州

【摘要】 目的探讨自体骨髓干细胞移植是否能增加缺血肢体的新生血管形成。方法抽取5只犬骨髓20ml,经Ficoll分离单核细胞,免疫磁珠分离CD34+细胞,在体外经血管内皮细胞生长因子诱导分化为内皮细胞,并经vWF抗染色鉴定。建立犬双侧肢体缺血动物模型,将Dil荧光标记的内皮细胞植入缺血肢体中。结果CD34+干细胞移植4周后,激光共聚焦显微镜证实缺血肢体中移植的自体骨髓干细胞参与了新生血管形成;动脉造影见干细胞移植的肢体侧枝循环明显增加,微血管密度检测移植组也明显高于对照组(12.0±2.8vs.5.0±1.6/每高倍镜视野,t=4.17,P<0.05);干细胞移植的肢体ABI平均为0.58±0.14,对照肢体ABI平均为0.32±0.11,差异有统计学意义(t=2.95,P<0.05)。结论自体骨髓干细胞移植能有效的促进缺血组织内的新生血管形成。

【Abstract】 Objective To explore whether transplantation of autologous bone marrow stem cells might augment angiogenesis and collateral vessel formation in a canine model of hindlimb ischemia. Methods CD34+ stem cells were centrifugation through Ficoll and an immune magnetic cell sorting system from bone marrow(20 ml) of canine (n=5) and induced into endothelial cells with VEGF in vitro, and expression of von Willebrand factor. Bilateral hindlimb ischemia was surgically induced in canines and Dil fluorescence labeled autologous stem cells were transplanted into the ischemic tissues. Results Four weeks after transplantation, fluorescence microscopy revealed that transplanted cells were incorporated into the capillary network among preserved skeletal myocytes. The stem cells transplanted group had more angiographically detectable collateral vessels, a higher capillary density (12.0±2.8 vs. 5.0±1.6 per field; t=4.17 P<0.05) and a higher ABI (0.58±0.14 vs. 0.32±0.11; t=2.95, P<0.05). Conclusions Direct local transplantation of autologous bone marrow CD34+ stem cells seems to be a useful strategy for therapeutic neovascularization in ischemic tissues in adults, consistent with “therapeutic vasculogenesis.”

【关键词】 骨髓移植局部缺血血管生成
【Key words】 Bone marrowTransplantationIschemiaAngiogenesis
【基金】 国家自然科学基金资助项目(30170918,30300335)
  • 【文献出处】 中华外科杂志 ,Chinese Journal of Surgery , 编辑部邮箱 ,2005年19期
  • 【分类号】R658
  • 【被引频次】15
  • 【下载频次】168
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