脂肪组织中血管基质部分细胞向内皮细胞诱导分化的实验研究  

The Experiment of Inducing Stromal Vascular Fraction (SVF) Cells Derived from Human Adipose into Endothelial Cells

作　　者：刘波[1] 朱金土[1] 曹毅[1] 高寿松[1] 余土根[1] 尹烁[2] 刘广鹏[2] 崔磊[2] 曹谊林[2] 

机构地区：[1]浙江省中医院整形外科,浙江省杭州市310006 [2]上海组织工程研究与开发中心,200235

出　　处：《组织工程与重建外科杂志》2008年第1期6-8,共3页Journal of Tissue Engineering and Reconstructive Surgery

摘　　要：目的探讨脂肪组织中血管基质部分(Stromal vascular fraction,SVF)细胞向内皮细胞诱导分化的方法。方法脂肪抽吸术获取人脂肪组织,消化法得到的细胞接种在FN包被的培养皿内,细胞培养,传至第2代,分诱导组和非诱导组进行培养。免疫细胞荧光分别检测诱导组和非诱导组的vWF表达;流式细胞仪分别检测诱导组和非诱导组的CD34、CD45、CD133和PECAM-1表达率;荧光显微镜观察细胞摄取DiI-ac-LDL的功能。结果诱导组细胞12d后呈现内皮细胞典型的铺路石样形态;免疫细胞荧光显示诱导组vWF表达阳性;荧光显微镜观察显示诱导组细胞具有摄取DiI-ac-LDL的功能;流式细胞仪检测显示诱导组PECAM-1阳性率为(67.41±13.35)%,明显高于非诱导组的(6.73±2.21)%(p<0.01),非诱导组CD34阳性率(72.39±13.45)%,明显高于诱导组的(16.06±3.86)%阳性率(p<0.01)。结论脂肪组织中血管基质部分细胞能够诱导分化为成熟内皮细胞,有望为血管组织工程提供新的种子细胞来源。Objective To investigate the methods of inducing SVF cells derived from human adipose into endothelial cells. Methods The cells obtained from human liposuction were plated on culture dishes coated with human fibronectin and were cultured in DMEM containing 2 % FBS. Cells of passage 2 cultured in EGM-2 （2 % FBS） served as the induced cells （experimental group）, with cells cultured in DMEM （2 % FBS） as the non-induced cells （control group）. Immunofluorescence was used to detect the expression of vWF. The function of taking up DiI-ac-LDL by the induced cells was observed through Fluorescence microscope. FACS was used to quantitatively analyze the expression rate of cell markers （CD34,CD45 ,CD133 and PECAM-1）. Results The induced cells of passage 2 exhibited cobblestone morphology, similar to that of the endothelial cells. vWF was positive in induced cells. Fluorescence microscopy observed the induced cells took up low-density lipoprotein （LDL）. FACS analysis showed （67.41 ± 13.35） % of the induced cells expressed PECAM-1 and （6.73 ± 2.21） % of the non-induced cells expressed PECAM-1 （p〈0.01）, while （72.39 ± 13.45） % of the non-induced cells expressed CD34 and （16.06 ± 3.86） % of the induced cells expressed CD34 （p〈0.01）. Conclusion SVF cells derived from human adipose may serve as another source of seeding cells for vascular tissue engineering.

关 键 词：人脂肪组织 血管基质部分细胞 内皮细胞 组织工程 

分 类 号：Q813.11[生物学—生物工程]