【摘要】 目的:探讨大鼠骨髓来源的血管内皮祖细胞(endothelial progenitor cells,EPCs)移植促进缺血皮瓣的血管新生,从而提高皮瓣存活率的可能性。方法:在Wistar大鼠背部形成2cm×8cm蒂部位于双侧髂棘连线上的随意型超比例皮瓣,建立缺血皮瓣模型。采用密度梯度离心法从Wistar大鼠骨髓中分离出骨髓单个核细胞,并在专有的诱导培养基(EG-M2MV)中培养,贴壁筛选法分离,诱导分化为EPCs;对细胞的形态、表型、功能加以鉴定。将EPCs注射移植于iWstar大鼠背部的缺血皮瓣,大体观察皮瓣的存活率,vWF免疫酶染色法检测皮瓣毛细血管密度。以缺血皮瓣单纯注射磷酸盐缓冲液(PBS)的Wistar大鼠作为对照组。结果:骨髓中分离培养的EPCs呈现典型的"铺路石"样外观,表达CD34、Flk-1及CDl 33表型,能够摄取DIL-ac-LDL和特异性结合FITC-UEA-1。注射EPCs组的缺血皮瓣存活率以及毛细血管密度显著高于对照组(P<0.05)。结论:骨髓中的EPCs在体外培养后注射移植于皮瓣内,可以促进缺血性皮瓣的血管新生,提高缺血性皮瓣的存活率。更多还原

【Abstract】 0bjective To discuss the neovascularization of transplanted rats bone marrow derived endothelial progenitor cells to ischemic flap,also it increased the survival rates of ischemia flaps.Methods Wistar rat model with topical ischemia skin flap was established by forming 2cm×8cm skin flap on the rat back.The marrow mononuclear cells were gained from Wistar rat bone marrow by density gradient centrifugation,and cultured in microvascular endothelial growth medium-2.Cells were isolated with adherence screening method and induced to differentiate to EPCs.EPCs were identified by cellular morphology,cellular phaenotype and cellular function.Then EPCs were injected into different ischemia skin flap sites.Gross observation of the survival rate of the ischemia flap and capillary vessel density evaluation with vWF immunoenzymatic staining were conducted.Wistar rats with PBS injection in ischemia skin flap were treated as control groups.Results Bone marrow derived EPCs formed typical cobblestone-like structure and expressed cell markers CD34、Flk-1 and CDl33.They took DIL-ac-LDL and specific binding FITC-UEA-1.The survival rate and capillary density of EPCs injection group were significantly higher than than the control group(P<0.05).Conclusion EPCs in bone marrow after vitro culture injected in transplated flaps,could improving the neovascularization of ischemia skin flaps and was increasing the survival rates of ischemia skin flaps.更多还原