人脂肪干细胞向血管平滑肌细胞诱导分化实验研究.docVIP

人脂肪干细胞向血管平滑肌细胞诱导分化实验研究 　　[摘要]目的:体外诱导培养人脂肪干细胞(adipose-derived stem cells, ADSCs),观察是否可以分化形成血管平滑肌细胞(vascular smooth muscle cells, VSMCs),为构建组织工程化血管寻找新的种子细胞来源。方法:免疫磁珠法分选收集原代脂肪干细胞,加入PDGF-BB、TGF-β1诱导14天后进行检测。结果:实验组细胞生长呈现血管平滑肌细胞所特有的峰谷样生长,血管平滑肌细胞特有的表面抗原标记物表达阳性。结论:脂肪干细胞定向诱导培养后,具有血管平滑肌细胞的特性,有可能成为构建组织工程化血管新的种子细胞来源。 　　[关键词]脂肪干细胞;血管平滑肌细胞;血小板衍生生长因子;转化生长因子β1 　　[中图分类号]Q813.1 R622.4 [文献标识码]A [文章编号]1008-6455(2010)02-0215-03 　　 　　Experiments of adscs induced into vascular smooth muscle cells 　　JIANG Wei1,CAO Qiang1,WANG Ji-hua2,FENG Xing-hua1 　　(1.Department of Oral and Maxillofacial Surgery,School of Stomatology,Fourth Military Medical University,Xian 710032,Shaanxi,China; 2. Department of Plastic Surgery,the Second Affiliated Hospital of Kunming Medical College, Kunming 650101,Yunnan,China) 　　 　　Abstract:ObjectiveTo investigate the feasibility of ADSCs differentiating into VSMCs in vitro and to find the new source of seeding cells in constructing the tissue-engineered vessels.MethodsTo separate the cells from the original adipose stem cells by MACS and culture 14 days with PDGF-BB and TGF-β1.ResultsThe cells show peak and valley growth pattern specified as mature VSMCs and express the VSMCs markers.ConclusionADSCs were induced in vitro in specific environment,the cells show the significant characteristics of VSMCs and have the potentiality being seeding cells of construct the tissue-engineered vessels. 　　Key words: adipose-derived stem cells; vascular smooth muscle cells; platelet derived growth factor BB; transforming growth factor beta 　　 　　脂肪干细胞由中胚层分化发育而来,具有类似于骨髓间充质干细胞的干细胞特性[1],可以向脂肪细胞、软骨细胞、肌肉细胞和成骨细胞分化[2],而且脂肪干细胞取材更方便、更容易获取。研究发现:体外培养ADSCs可表达肌源性标志物MyoD1 和肌球蛋白重链[3],体内实验也证明其表达α- 肌动蛋白[4]。本实验观察研究了脂肪干细胞在特定的诱导环境下,向血管平滑肌细胞分化的情况。 　　 　　1材料和方法 　　 　　1.1 主要试剂与仪器:FBS、M-199 培养基(Gibco公司,美国);TGF-β1(transforming growth factor, TGF-β1)、PDGF-BB(platelet derives growth factor-BB, PDGF-BB)(Sigma公司,美国);单克隆鼠抗人α平滑肌肌动蛋白(α-SMA,Dako Cytomation公司,美国);单克隆鼠抗人平滑肌肌球蛋白重链(SM-MHC,Chemicon公司,加拿大);单