Tang Ting-ting, Xu Xiao-liang, Dai Ke-rong, Yu Chao-feng, Yue Bing, Lou Jue-ren
Bone and Joint Research Center, Department of Orthopaedic Surgery, Shanghai Ninth People's Hospital, Shanghai 200011, China.
Chin J Traumatol. 2005 Feb;8(1):3-7.
To evaluate the osteogenic potential of bone morphogenetic protein (BMP)-2 gene transfected goat bone marrow-derived mesenchymal stem cells (MSCs).
Goat bone marrow-derived MSCs were transfected by Adv-human bone morphogenetic protein (hBMP)-2 gene (Group 1), Adv-beta gal transfected MSCs (Group 2) and uninfected MSCs (Group 3). Western blot analysis, alkaline phosphatase staining, Von Kossa staining and transmission electron microscopy were adopted to determine the phenotype of MSCs. Then the cells were injected into thigh muscles of the nude mice. Radiographical and histological evaluations were performed at different intervals.
Only Adv-hBMP-2 transfected MSCs produced hBMP-2. These cells were positive for alkaline phosphatase staining at the 12th day and were positive for Von Kossa staining at the 16th day after gene transfer. Electron microscopic observation showed that there were more rough endoplasmic reticulum, mitochondria and lysosomes in Adv-hBMP-2 transfected MSCs compared to MSCs of other two groups. At the 3rd and 6th weeks after cell injection, ectopic bones were observed in muscles of nude mice of Group 1. Only fibrous tissue or a little bone was found in other two groups.
BMP-2 gene transfected MSCs can differentiate into osteoblasts in vitro and induce bone formation in vivo.
评估骨形态发生蛋白(BMP)-2基因转染的山羊骨髓间充质干细胞(MSCs)的成骨潜能。
用腺病毒介导的人骨形态发生蛋白(hBMP)-2基因转染山羊骨髓间充质干细胞(第1组),用腺病毒介导的β半乳糖苷酶转染间充质干细胞(第2组),未感染的间充质干细胞作为对照(第3组)。采用蛋白质免疫印迹分析、碱性磷酸酶染色、Von Kossa染色及透射电镜观察来确定间充质干细胞的表型。然后将细胞注射到裸鼠大腿肌肉中。在不同时间间隔进行影像学和组织学评估。
仅腺病毒介导的hBMP-2转染的间充质干细胞产生hBMP-2。基因转染后第12天这些细胞碱性磷酸酶染色呈阳性,第16天Von Kossa染色呈阳性。电镜观察显示,与其他两组间充质干细胞相比,腺病毒介导的hBMP-2转染的间充质干细胞有更多的粗面内质网、线粒体和溶酶体。细胞注射后第3周和第6周,在第1组裸鼠肌肉中观察到异位骨形成。其他两组仅发现纤维组织或少量骨组织。
BMP-2基因转染的间充质干细胞在体外可分化为成骨细胞,并在体内诱导骨形成。