Yi C, Zheng Q, Guo X, Liu Y
Department of Orthopedics, Xiehe Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022.
J Tongji Med Univ. 2001;21(2):130-3. doi: 10.1007/BF02888076.
To study the osteogenic potential of cultured bone marrow stromal cells (BMSCs) transfected with transforming growth factor beta 1 (TGF-beta 1) gene in vitro, cultured BMSCs were transfected with the complexes of pcDNA3-TGF-beta 1 and Lipofectamine Reagent in vitro. The cell proliferation was detected by MTT method and the morphological features of transfected BMSCs was observed. ALP stains and PNP method were used to measure ALP activity. In addition, the collagen type I propeptides and mineralized matrixes were examined by immunohistochemical staining and tetracycline fluorescence labeling respectively. The morphological and biological characters of the transfected BMSCs were similar to those of osteoblasts and the cell proliferation was promoted. The cell layer displayed strong positive reaction for ALP stains and immunohistochemical staining. ALP activity and collagen type I expression increased remarkably after transfection. Mineralized matrixes formed earlier and more in transfected BMSCs as compared with control group. It is concluded that transfecting with TGF-beta 1 gene could promote the osteogenic potential of cultured BMSCs.
为研究体外转染转化生长因子β1(TGF-β1)基因的培养骨髓基质细胞(BMSCs)的成骨潜能,体外将pcDNA3-TGF-β1与脂质体转染试剂的复合物转染培养的BMSCs。采用MTT法检测细胞增殖情况,并观察转染后BMSCs的形态学特征。运用碱性磷酸酶(ALP)染色和对硝基苯酚(PNP)法检测ALP活性。此外,分别通过免疫组织化学染色和四环素荧光标记检测Ⅰ型胶原蛋白前肽和矿化基质。转染后的BMSCs的形态学和生物学特性与成骨细胞相似,且细胞增殖得到促进。细胞层对ALP染色和免疫组织化学染色呈强阳性反应。转染后ALP活性和Ⅰ型胶原蛋白表达显著增加。与对照组相比,转染后的BMSCs中矿化基质形成更早且更多。结论是,转染TGF-β1基因可促进培养的BMSCs的成骨潜能。