血管内皮细胞生长因子基因转染血管内皮祖细胞移植提高移植脂肪存活率的实验研究  

Experimental study of transplanted endothelial progenitor cells transfected with VEGF165 gene augment the survival volume of transplanted fat tissue

作　　者：易成刚[1] 郭树忠[1] 张琳西[1] 夏炜[1] 韩岩[1] 舒茂国[1] 张辉[1] 周庆红[1] 

机构地区：[1]第四军医大学西京医院整形外科中心,西安710032

出　　处：《中华外科杂志》2005年第11期730-735,共6页Chinese Journal of Surgery

基　　金：国家自然科学基金资助项目(30400471)

摘　　要：目的探讨血管内皮细胞生长因子165(VEGF165)基因转染血管内皮祖细胞(EPCs)移植促进游离移植的脂肪组织的血管新生,提高移植脂肪组织存活率。方法体外分离、培养人脐血中EPCs,利用脂质体介导VEGF165基因体外转染EPCs,然后与来自人体的脂肪组织混合移植于裸鼠背部,裸鼠随机分为3组:VEGF165基因转染组、EPCs组及M199培养基对照组。结果脐血中分离培养的EPCs表达CD34、血管内皮细胞生长因子受体及CD133;VEGF165基因转染EPCs体外及体内检测均有VEGF165蛋白的表达。VEGF165基因转染组、EPCs组中,EPCs整合到缺血部位新生血管中,与对照组的脂肪组织存活率分别为(96.2±8.6)%、(75.3±6.8)%和(40.2±2.5)%(P<0.05),VEGF165基因转染组与EPCs组脂肪组织周边区毛细血管密度有显著差异(P<0.05),均高于对照组(P<0.05)。术后3个月时3组脂肪组织周边区的EPCs密度分别为(196±16)个/mm2、(95±11)个/mm2、0个/mm2(P<0.05)。结论脐血中的EPCs体外培养后移植体内可促进游离移植的脂肪组织的血管新生,提高存活率,而转染VEGF165基因的EPCs具有更强的促血管新生的作用。Objective To investigate the feasibility of transplanting endothelial progenitor cells(EPCs) transfected with VEGF165 gene to free transplanted fat tissue for increasing neovascularization and the survival. Methods EPCs isolated from human cord blood were cultured in vitro and identified by immunocytochemistry. After transfection by VEGF165 gene, the expression of VEGF was assessed using ELISA. Then EPCs with (VEGF gene transfection group) and without VEGF165 gene transfection (EPCs group) were transplanted to free transplanted fat tissue at 18 nude mice's back, and nine nude mice transplanted with free fat tissue were injected with M199 (control group). CM-DiI was used to trace the transplanted cells. The capillary density of transplanted fat tissue was detected by CD34 immunohistochemistry. Results EPCs expressed cell markers CD34、KDR and CD133. After transfection, the expression of VEGF was positive. Transplanted EPCs survived and proliferated, and transplanted EPCs were incorporated into the capillary networks in the transplanted fat tissue. The percent of survival volume of transplanted fat tissue of VEGF gene transfection group was (96.2±8.6)%, significantly higher than that of the EPCs group [(75.3±6.8)%, P<0.05) and M199 group [(40.2±2.5)%, P<0.05). The capillary density of transplanted fat tissue of VEGF gene transfection group was significantly higher than those of the EPCs group and M199 group (P<0.05). Conclusions EPCs from human cord blood can increase free transplanted fat tissue neovascularization and the survival volume, and the ability of promoting neovascularization of EPCs transfected with VEGF165 gene is more potent than EPCs alone.

关 键 词：基因转染 存活率 祖细胞移植 血管内皮祖细胞(EPCs) 实验研究 VEGF165基因 血管内皮细胞生长因子受体 脂肪组织 毛细血管密度 游离移植 脂质体介导 CD133 促血管新生 对照组 体外分离 体外转染 混合移植 CD34 分离培养 

分 类 号：R622.9[医药卫生—整形外科]