脂肪干细胞及血管内皮细胞提高游离脂肪移植成活的实验研究  

Effects of adipose-derived stem cells and endothelial cells on survival and neovascularization of fat tissue transplants

作　　者：张俊慧 熊炳钧 邱娟娟 谭秋雯 张忆[2] 杨济桥[1] 吕青[1] ZHANG Junhui;XIONG Bingjun;QIU Juanjuan;TAN Qiuwen;ZHANG Yi;YANG Jiqiao;LU Qing(Department of Breast Surgery,West China Hospital,Sichuan University,Chengdu Sichuan,610041,P.R.China;Division of Stem Cell and Tissue Engineering,State Key Laboratory of Biotherapy,West China Hospital,Sichuan University,Chengdu Sichuan,610041,P.R.China)

机构地区：[1]四川大学华西医院乳腺外科,成都610041 [2]四川大学华西医院生物治疗国家重点实验室干细胞与组织工程研究室,成都610041

出　　处：《中国修复重建外科杂志》2018年第8期1074-1080,共7页Chinese Journal of Reparative and Reconstructive Surgery

摘　　要：目的探讨脂肪来源干细胞(adipose-derived stem cells,ADSCs)及血管内皮细胞(endothelial cells,ECs)对脂肪移植术后血管新生及脂肪成活的影响。方法取乳房切除术患者自愿捐赠的脂肪组织,采用胶原酶消化法分离培养ADSCs,取第3代细胞进行实验;同时制备游离脂肪颗粒。取健康4~6周龄雄性裸鼠80只,随机分为4组(n=20),分别于裸鼠背部皮下注射1 m L脂肪颗粒+0.3 m L生理盐水(对照组)、1 m L脂肪颗粒+2×106个人脐静脉内皮细胞株(human umbilical vein endothelial cells,HUVECs)+0.3 m L生理盐水(ECs组)、1 m L脂肪颗粒+2×10!6个ADSCs+0.3 m L生理盐水(ADSCs组)、1 m L脂肪颗粒+1×10~6个HUVECs+1×10!6个ADSCs+0.3 m L生理盐水(ADSCs+ECs组)。注射后观察各组裸鼠存活情况,大体观察背部移植区色泽、形状。2、4、8、12周时背部移植区行超声影像学观察,取标本采用排水法测量其体积,组织学观察移植脂肪大体结构变化情况,免疫荧光染色观察血管新生情况。结果各组裸鼠均存活至实验完成。移植后各时间点各组超声检测结果无明显差异,移植脂肪内部均未见明显血流信号,未见囊肿、钙化、实性占位等异常表现。移植后各时间点,ADSCs组及ADSCs+ECs组移植脂肪体积均高于ECs组及对照组(P<0.05);且8、12周时ADSCs组明显高于ADSCs+ECs组(P<0.05)。HE染色示移植后各组组织学形态变化趋势相似,但ADSCs组组织重塑速度快于其他各组。免疫荧光染色示随时间延长各组新生血管逐渐增加,各时间点ECs组、ADSCs组、ADSCs+ECs组微血管密度均高于对照组(P<0.05);4周时ADSCs组微血管密度高于ECs组及ADSCs+ECs组(P<0.05);8、12周时ADSCs组及ADSCs+ECs组高于ECs组(P<0.05),ADSCs组及ADSCs+ECs组间比较差异无统计学意义(P>0.05)。结论 ADSCs可提高移植脂肪存活率,且该促进作用可能与促进血管新生相关。Objective To investigate the effects of adipose-derived stem cells（ADSCs） and endothelial cells（ECs） on the survival and neovascularization of fat tissue transplants. Methods The ADSCs were isolated by collagenase digestion from the adipose tissues voluntarily donated by the patients undergoing mastectomy, and subcultured. The passage 3 ADSCs were used for subsequent experiments. The residual fat tissues were used to prepare fat particles（FPs）.The human umbilical vein endothelial cells（HUVECs） were used as ECs for subsequent experiments. Eighty healthy male nude mice, aged 4-6 weeks, were randomly divided into 4 groups（n=20）. The mice were received subcutaneous injection at the dorsum of 1 m L FPs＋0.3 m L normal saline（NS） in control group, 1 m L FPs＋2×106 ECs＋0.3 m L NS in ECs group,1 m L FPs＋2×10~6 ADSCs＋0.3 m L NS in ADSCs group, and 1 m L FPs＋1×10~6 ECs＋1×10~6 ADSCs＋0.3 NS in ADSCs＋ECs group. General observations of the injection sites were performed, and the survival of the mice was recorded. At 2, 4, 8,and 12 weeks after injection, grafted fat tissues were firstly assessed by ultrasonography, then they were collected for volume measurement（water displacement method） and histology observation（HE staining and immunofluorescence staining）. Results All mice survived until the end of experiment. At each time point, no significant difference was noted between groups in ultrasonography assay. There was no significant blood flow signal in the grafted fat tissues, or cysts,calcification, solid occupying in recipient area. Generally, the volume of grafted fat tissues decreased with time in all groups. Specifically, the volumes of grafted fat tissues were larger in ADSCs group and ADSCs＋ECs group than that in control group and ECs group（P〈0.05） at each time point, and in ADSCs group than in ADSCs＋ECs group（P〈0.05） at8 and 12 weeks. HE staining showed that all groups had similar tendencies in general histology changes, and remodeling in ADSCs group w

关 键 词：脂肪移植 血管内皮细胞 脂肪来源干细胞 裸鼠 

分 类 号：R622.9[医药卫生—整形外科]