三种不同颗粒脂肪来源干细胞增殖、分化、迁移能力的比较  

Comparison of proliferation, differentiation and migration ability of adipose-derived stem cells in three different granular fats

作　　者：廖选[1] 陈嘉文 黄苏 梁俊贤 李升红[1] 肖丽玲[1] 刘宏伟[1] Liao Xuan;Chen Jiawen;Huang Su;Liang Junxian;Li Shenghong;Xiao Liling;Liu Hongwei(Department of Plastic Surgery,the First Affiliated Hospital of Jinan University,Innovative Technology Research Institute of Tissue Repair and Regeneration,Key Laboratory of Regenerative Medicine,Ministry of Education,Guangzhou 510630,China)

机构地区：[1]暨南大学附属第一医院整形外科,暨南大学整形外科新技术研究所,再生医学教育部重点实验室,广州510630

出　　处：《中华整形外科杂志》2019年第8期796-803,共8页Chinese Journal of Plastic Surgery

基　　金：国家自然科学基金(81871563);广州市科技计划资助项目(201508020253,201300000091);暨南大学第一临床医学院科研培育专项基金(2017306);中央高校基本科研业务费专项资金(21619350)

摘　　要：目的比较3种不同颗粒脂肪来源干细胞增殖、分化及迁移的能力。方法收集2016年6月至2019年6月10例就诊于暨南大学附属第一医院行髂腰部脂肪抽吸术患者的废弃脂肪组织,均为女性,年龄20~35岁,利用不同直径的抽脂针和不同大小侧孔的脂肪抽吸器械分别各获取10 ml颗粒脂肪(n=10)、10 ml微粒脂肪(n=10)及10 ml超微粒脂肪(n=10)。酶消化法分离提取脂肪来源干细胞(adipose-derived stem cells,ADSCs),分析3种脂肪来源血管基质成分的差异,XTT法检测其增殖生长能力,体外细胞创伤模型观察其迁移能力,成脂、成骨诱导实验分析比较其多向分化能力。实验数据以SPSS 13.0软件进行分析,多组样本均值比较应用One-Way ANOVA,P<0.05为差异具有统计学意义。结果流式细胞仪检测颗粒脂肪、微粒脂肪、超微粒脂肪中ADSCs含量分别为(4.23±0.37)×10^5、(2.29±0.33)×10^5和(1.56±0.16)×10^5,颗粒脂肪中含量最高,其次为微粒脂肪,超微粒脂肪中最少,差异有统计学意义(F=209.533,P<0.001)。XTT法结果示第1和第2代颗粒脂肪中ADSCs增殖能力最高,其次为微粒脂肪,而超微粒脂肪的增殖能力最低,3组间吸光度值呈时间依赖性增长;第3代3组细胞增殖能力无明显差异。细胞创伤模型显示第1代颗粒脂肪ADSCs迁移能力最好,其次为微粒脂肪,而超微粒的迁移能力最弱,3组间0、12、24 h创面剩余面积比较,F=306.370、1409.907,P值均<0.001;第2代起各组ADSCs迁移能力逐渐自我修复,12 h迁移能力与第1代相似(F=11.410,P<0.001),而24 h迁移能力则无明显差异(F=0.070,P=0.933)。油红O及茜素红染色显示第1代颗粒脂肪和微粒脂肪的成脂、成骨分化能力比超微粒脂肪强(F=523.532、620.022,P值均<0.001),第2代起则无明显变化(F=2.144、0.866,P=0.137、0.432)。结论超微粒脂肪在脂肪组织抽取的过程中,物理切割等因素会对ADSCs的产量及活性造成影响。而在培养、传代的过程中,�Objective To study the effects of three different granular fats on proliferation,differentiation and migration of adipose-derived stem cells.Methods Ten patients were selected for lumbar liposuction.The adipose tissue was obtained with different sized side-hole fat aspiration devices:10 ml of Macrofat(n=10),10 ml of Microfat(n=10)and 10 ml of Nanofat(n=10).Enzyme digestion method was used to separate and extract adipose-derived stem cells(ADSCs).The differences of vascular matrix components in these 3 kinds of fat sources were analyzed.XTT assay was used to detect proliferation and growth ability.The migration ability of the cell injury model was observed in vitro,and the multi-directional differentiation ability was compared by the analysis of adipogenic and osteogenic induction.The experimental data were analyzed using SPSS 13.0 software.One-Way ANOVA was used to compare the difference of multiple groups.P<0.05 was considered as statistically significant.Results The detection of adipose-derived stem cells was by flow cytometry and it showed Macrofat,Microfat and Nanofat was(4.23±0.37)×10^5,(2.29±0.33)×10^5 and(1.56±0.16)×10^5.The content of fat-derived stem cells in Macrofat was the highest,followed by Microfat,and the content of Nanofat was the least(F=209.533,P<0.001).XTT assay showed that the proliferative ability of adipose-derived stem cells in the first two generations of Macrofat was the highest,followed by Microfat,while the proliferation of Nanofat was the lowest(the absorbance in 3 groups increased in a time-dependent manner).There was no significant difference in the proliferative capacity of the third generation of 3 groups(the absorbance of these 3 groups were compared at each time point).The cellular trauma model showed that the first generation of Macrofat-derived stem cells had the best migration ability,followed by Microfat,while the Nanofat had the weakest migration ability(Compared with the remaining area of wounds at 0 h,12 h,24 h point between the three groups,F=306.370,1409.907,P<0.0

关 键 词：脂肪颗粒 超微粒脂肪 脂肪来源干细胞 增殖 分化 迁移 

分 类 号：R32[医药卫生—人体解剖和组织胚胎学]