人脂肪干细胞与表皮生长因子促进皮肤创面愈合的研究  

Human adiposederived stem cells and epidermal growth factor promoting skin wound healing

作　　者：张春连 孙振龙[1] 王璐[2] Zhang Chunlian;Sun Zhenlong;Wang Lu(Department of Plastic Surgery,Zhengzhou Third People′s Hospital,Zhengzhou 450016,China;Department of Plastic Surgery,the Second Affiliated Hospital of Zhengzhou University,Zhengzhou 450014,China)

机构地区：[1]郑州市第三人民医院整形外科,450016 [2]郑州大学第二附属医院整形外科,450014

出　　处：《中华实验外科杂志》2021年第9期1690-1695,共6页Chinese Journal of Experimental Surgery

摘　　要：目的观察人脂肪干细胞(hASCs)与人表皮生长因子(hEGF)对皮肤缺损创面修复的影响。方法选择郑州大学第二附属医院2020年1月6例患者抽脂减肥的脂肪组织作为hASCs的来源,以酶消化法从人脂肪组织中提取hASCs,将其培养至第3代。利用倒置显微镜对细胞形态进行观察;使用流式细胞仪鉴定细胞表型及分化能力检测。随机将24只新西兰大白兔建立背部全层皮肤缺损模型,并将实验用新西兰大白兔随机分为3组,设立实验组(hASCs+hEGF组),细胞治疗对照组(hASCs组)和空白对照组[磷酸盐缓冲液(PBS)组]。实验组予以100 μl PBS重悬第3代脂肪来源干细胞(1×106个),以10 μg/L加入人表皮细胞生长因子;细胞治疗对照组予以100 μl PBS重悬第3代脂肪来源干细胞(1×106个);空白对照组取PBS,各组均注射600 μl局部移植至创面边缘及基底部。大体观察创面愈合情况,计算创面面积占初始创面面积百分比;创面组织进行标本苏木精-伊红(HE)染色,免疫组织化学CD31染色检测等方法观察和比较各组创面愈合水平。多样本均数比较采用方差分析,采用t检验对比组间计量资料。结果相差显微镜观察hASCs的细胞形态学特征:可见原代脂肪间充质干细胞(ADSCs)呈梭形,接种后2 h后开始圆形,大部分细胞变形在48 h后,细胞形态以长梭形为主,短梭形、狭长形及多角形较少,胞质丰富,胞核清楚,并分泌大量基质,基质内呈特异性深蓝色,第3代时可见细胞以长梭形为主,漩涡状生长。应用流式细胞仪检测显示CD90、CD105、CD73呈阳性表达,阳性率在95%以上,CD34、CD11b、CD19、CD45及HLADR呈阴性表达;人脂肪源性干细胞(hADSCs)在加入成脂诱导液培养后,hADSCs可向脂肪细胞分化,加入成软骨诱导培养基诱导后,hADSCs可向软骨细胞分化。表明提取培养的细胞为hADSCs。实验组创面完全愈合,被覆盖新生的上皮组织,接近正常皮肤;细胞治疗对照组创面Objective To observe the effects of human adipose-derived stem cells(hASCs)and human epidermal growth factor(hEGF)on the repair of defects.Methods Adipose tissue from 6 patients with liposuction and weight loss in the Second Affiliated Hospital of Zhengzhou University in January 2020 was selected as the source of human adipose stem cells,and hASCs were extracted from human adipose tissue by enzymatic digestion and cultured to the third generation.The cell morphology was observed by an inverted microscope,and phenotypes were identified by flow cytometry,and differentiation ability was detected.Twenty-four New Zealand white rabbits were used to establish a full-thickness skin defect model of the back and were randomly divided into three groups:experimental group(hASCs+hEGF),cell therapy control group(hASCs),and blank control group(PBS).The experimental group was transplanted to the edge and the base of the wound by injection of 600μl that the passage 3 adipose-derived stem cells(1×106)were suspended through 100μl PBS was added with human epidermal growth factor for 10μg/L.The cell therapy control group was transplanted to the edge and the base of the wound by injection of 600μl that the passage 3 adipose-derived stem cells(1×106)were suspended through 100μl PBS.The blank control group was transplanted to the edge and the base of the wound by injection of 600μl PBS.They were used to observe and compare the wound healing level of each group that the wound healing was observed and the percentage of wound area in the initial wound area was calculated,and hematoxylin and eosin(HE)staining and CD31 immunohistochemical staining were used to detect the wound tissue.The analysis of variance was used to compare the means of multiple samples,and the T-test was used to compare the measurement data between groups.Results The morphological characteristics of hASCs were observed by a phase-contrast microscope:the primary ADSCs were spindle-shaped and began to be round after 2 hours of inoculation.Most of the cells were de

关 键 词：人脂肪干细胞 表皮生长因子 创面 

分 类 号：R641[医药卫生—外科学]