同时分离和培养兔外周血平滑肌祖细胞和内皮祖细胞

同时分离和培养兔外周血平滑肌祖细胞(SPC)和内皮祖细胞(EPC),为组织工程膀胱的构建和血管化提供种子细胞.分离新西兰兔外周血中的单个核细胞,分别进行SPC和EPC的分离和培养;同时,培养兔膀胱平滑肌细胞(BSMC)作为对照.细胞爬片,观察细胞摄取Dil-AcLDL和结合FITC-UEA-1的能力;间接免疫荧光染色观察平滑肌肌动蛋白(SMA)、结蛋白(Desmin)、KDR、eNOS和vWF的表达情况;进行细胞增殖实验,观察PDGF-BB,VEGF对SPC和EPC的增殖作用.结果发现,培养1周后出现SPC克隆和EPC克隆,SPC呈梭形,长短不一;EPC形态均一,呈典型的鹅卵石形;培养的BSMC形态均一,为长梭形,呈峰谷样形态.利用克隆环分离SPC和EPC克隆,继续培养可得到高纯度的SPC和EPC.SPC不摄取Dil-AcLDL,不结合FITC-UEA-1;SPC表达SMA、Desmin和KDR,不表达eNOS和vWF.EPC同时摄取Dil-AcLDL和结合FITC-UEA-1;EPC表达eNOS、vWF和KDR,不表达SMA和Desmin.BSMC仅表达SMA和Desmin.PDGF-BB仅能促进SPC增殖...

Simultaneous isolation and culture of smooth muscle progenitor cell and endothelial progenitor cell from rabbit peripheral blood

The objective of this study was to simultaneously isolate and culture smooth muscle progenitor cell (SPC) and endothelial progenitor cell (EPC) from rabbit peripheral blood for use as cell sources in tissue engineering of bladder and vascularization. In this study,SPC and EPC were simultaneously isolated and cultured from the peripheral blood of New Zealand Rabbit,and rabbit bladder smooth muscle (BSMC) were cultured and used as smooth muscle cell positive control. The cells were cultured on coverslips and ...