表达人骨形成蛋白2的成纤维细胞系的建立与鉴定  

Establishment and identification of expression human bone morphogenetic protein2 fibroblasts

作　　者：王娟[1] 孙卫斌[2] 卢春[3] 唐桂霞[3] 

机构地区：[1]南京医科大学口腔医学院牙周科,江苏南京210029 [2]四川大学华西口腔医学院牙周科,四川成都610041 [3]南京医科大学微生物与免疫学系,江苏南京210029

出　　处：《南京医科大学学报（自然科学版）》2005年第6期395-397,F002,共4页Journal of Nanjing Medical University(Natural Sciences)

摘　　要：目的:建立能稳定表达BMP2的成纤维细胞系。方法:运用阳离子聚合物转染试剂,将含有人BMP2基因的真核表达载体pcDNA3.1-B2导入NIH3T3细胞,通过G418筛选获得阳性细胞克隆,RT-PCR、免疫组化和酶联免疫吸附试验(ELISA)检测BMP2基因的稳定转染和蛋白表达。结果:转染细胞内有BMP2mRNA的转录,胞内及胞外有BMP2蛋白的表达。结论:采用阳离子聚合物转染法可成功地将外源性hBMP2基因导入NIH3T3细胞,为进一步研究诱导牙周膜细胞骨化分化建立基础。Objective:To establish fibroblasts stably expressing human bone morphogenetic protein2. Methods:Eukaryonic expression vector(pcDNA3.1-B2) was transduced into NIH3T3 cells by using SofastTM, a positive compound transfection agent. The positive cell clones were selected with G418. The stable transfection and expression of BMP2 in the NIH3T3 cells were determined by RT-PCR, immunohistochemical stain and Enzyme-linked immunosorbent assay(ELISA). Results:BMP2 mRNA was transcripted and expressed in the transfected NIH3T3 cells. Conclusion:With positive compound transfection, outside human BMP2 gene can be successfully transducted into NIH3T3 cells, which is the key step to induce periodontal cells to osseous phenotypes.

关 键 词：骨形成蛋白2 基因转染 成纤维细胞 

分 类 号：R322.71[医药卫生—人体解剖和组织胚胎学]