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

作　　者：张鸿坤[1] 张楠[2] 吴丽花[1] 金炜[1] 李鸣[1] 封华[1] 赵海格[1] 陈旭东[1] 杨春虎[3] 

机构地区：[1]浙江大学医学院附属第一医院血管外科,杭州310003 [2]浙江大学医学院附属邵逸夫医院内分泌科 [3]浙江大学医学院生物电磁学省重点实验室

出　　处：《中华外科杂志》2005年第19期1275-1278,共4页Chinese Journal of Surgery

基　　金：国家自然科学基金资助项目(30170918;30300335)

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。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 isehemie 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） anda higherABI （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 “herapeutic vasculogenesis. ”

关 键 词：骨髓 移植 局部缺血 血管生成 自体骨髓干细胞移植 CD34^+细胞 细胞移植治疗 肢体缺血 血管内皮细胞生长因子 实验研 

分 类 号：R658[医药卫生—外科学]