CD54^+和CD54^-脂肪干细胞的成脂分化能力  

Adipogenic capacity of CD54^+/CD54^- adipose-derived stem cells

作　　者：李德全 梁至洁[2] 韦金儒[3] 黄海[2] 黄敏红[2] 池刚毅[4] 黎洪棉[4] Li De-quan Liang Zhi-jie Wei Jin-ru Huang Hai Huang Min-hong Chi Gang-yi Li Hong-mian(Department of Breast Surgery, Third Hospital of Nanchang, Nanchang 330009, Jiangxi Province, China Department of Hepatobiliary and Gland Surgery Department of Vasculocardiology, 4Department of Plastic and Aesthetic Surgery, the Fifth Affiliated Hospital of Guangxi Medical University, Nanning 530022, Guangxi Zhuang Autonomous Region, China)

机构地区：[1]南昌市第三医院乳腺肿瘤一科,江西省南昌市330009 [2]广西医科大学第五附属医院肝胆腺体外科,广西壮族自治区南宁市530022 [3]广西医科大学第五附属医院心血管内科,广西壮族自治区南宁市530022 [4]广西医科大学第五附属医院整形美容外科,广西壮族自治区南宁市530022

出　　处：《中国组织工程研究》2017年第17期2638-2643,共6页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金资助项目(81560316;81560358);广西科学研究与技术开发计划资助项目(桂科攻1598012-1);广西自然科学基金重点项目(2016GXNSFDA380016);南宁市科学研究与技术开发计划资助项目(20153089;zc20153002)~~

摘　　要：背景:研究表明,脂肪干细胞具有多向分化潜能,但是其向成熟脂肪细胞分化的效率仍较低,只有30%-40%。因此,如何提高脂肪干细胞的成脂分化能力是软组织再生研究过程中需要解决的关键科学问题。目的:观察兔脂肪干细胞表面标志物CD54的表达与其成脂分化能力的关系,并探讨同一诱导剂对CD54^+和CD54^-兔脂肪干细胞成脂分化的影响。方法:取新西兰大白兔(8-12周龄)腹股沟皮下脂肪垫3 mL,采用贴壁法体外分离培养兔脂肪干细胞并行多向分化诱导及细胞表型标志物鉴定。取第3代细胞经免疫磁珠分选得到CD54^+兔脂肪干细胞和CD54^-兔脂肪干细胞,经成脂诱导14d后行油红O染色,检测成熟脂肪细胞的密度及细胞内脂滴含量。RT-PCR检测脂肪干细胞成脂分化相关基因表达水平。结果与结论:(1)兔脂肪干细胞具有间充质干细胞特性,可向成熟脂肪细胞、成骨细胞和软骨细胞表型分化,细胞表面标志物CD29、CD44、CD49d、CD54、CD73、CD90和CD105表达均为阳性,而CD31、CD34和CD45表达阴性;(2)成脂诱导第14天,CD54^+兔脂肪干细胞分化为成熟脂肪细胞的密度和细胞内脂滴含量均高于CD54^-兔脂肪干细胞,差异均有显著性意义(P<0.05);(3)成脂诱导14 d后,CD54^+兔脂肪干细胞成脂相关基因ADD1、C/EBPα、PPARγ的mRNA表达水平明显高于CD54^-兔脂肪干细胞,差异均有显著性意义(P<0.05);(4)结果表明,CD54^+脂肪干细胞具有高成脂分化能力。BACKGROUND： Studies have shown that adipose-derived stem cells have pluripotent differentiation potential, but only 30%-40% of cells can differentiate into mature adipocytes with low adipogenic differentiation potential. Therefore, how to improve the adipogenic differentiation ability of adipose-derived stem cells is a key problem to be solved in the process of soft tissue regeneration. OBJECTIVE： To observe the relationship between the surface marker CD54 of rabbit adipose-derived stem cells and their adipogenic capacity, and to explore the adipogenic differentiation of CD54＋/CD54- adipose-derived stem cells underthe same induction. METHODS： We successfully isolated and cultured the adipose-derived stem cells from inguinal subcutaneous fat pads （3 ml） of New Zealand white rabbits, aged 8-12 weeks, which were induced into multi-differentiation and used to detectsurface markers. We sorted the passage 3 adipose-derived stem cells by immunomagnetic beads and divided into two categories including CD54＋ and CD54- adipose-derived stem cells. After 14 days of adipogenic induction, the cells in the two groups were subjected to oil red O staining and were compared by detecting the density of mature adipocytes and lipid droplet contenT.RESULTS AND CONCLUSION： The cultured adipose-derived stem cells possessed the characteristics of mesenchymal stem cells that could differentiate into mature adipocytes, osteoblasts and chondrocytes, with CD29, CD44, CD49d, CD54, CD73, CD90 and CD105 positive expression while CD31, CD34 and CD45 negative expression. Fourteen days after adipogenic induction, the density of mature adipocytes and the intracellular lipid droplet content in the CD54＋ group were significantly higher than those in the CD54- group （P 〈 0.05）. We also found that the mRNA expressions of PPARγ,ADD1, C/EBPα related to adipogenic differentiation in the CD54＋ group were significantly higher than those in the CD54- group （P 〈 0.05）. Taken together, CD54＋ adipose-derived stem cells have ex

关 键 词：干细胞 脂肪干细胞 细胞表面标志 成脂分化 细胞诱导 组织工程 国家自然科学基金 

分 类 号：R394.2[医药卫生—医学遗传学]