【作者】 吴迪；

【导师】 梅陵宣；

【作者基本信息】 安徽医科大学 ， 口腔临床医学， 2006， 硕士

【摘要】 骨保护素基因修饰的大鼠骨髓基质细胞系的培养、表达与鉴定 目的：培养经OPG基因修饰的大鼠骨髓基质细胞系并检测其表达能力，鉴定表达产物，为基因工程技术治疗牙周病提供物质基础。 方法：本研究采用密度梯度离心法从成年SD大鼠骨髓中分离获得骨髓基质细胞，利用已构建的pSecTag-opg真核分泌表达穿梭载体，用脂质体法将目的基因转染至大鼠骨髓基质细胞，并应用免疫组织化学和western blot方法证实转染的细胞表达骨保护素。 结果：分离培养出大鼠骨髓基质细胞，经脂质体法转染目的基因并建立了稳定转染的细胞系。经免疫组织化学和western blot方法检测，在转染OPG基因的大鼠骨髓基质细胞中有骨保护素表达。 结论：本实验成功培养出分泌表达OPG的大鼠骨髓基质细胞系。更多还原

【Abstract】 Objective: In order to understand the ability of expression and secretion ofeukaryotic plasmid of osteoprotegerin, we transfected mammalian secreted expressive pSecTag2/B-OPG vector into rat MSC and detected its expression in rat MSC cell lines in this study, to prepare for gene engineering therapy of periodontitis.Method: Rat MSC cells were isolated by a density gradient and cultured from adultSD rat. The constructed recombinant plasmid was transfected into rat MSC cell line and OPG expressed in rat MSC cell line was identified by immunohistochemistry and western blot analysis.Result: Rat MSC cells were separated and cultured. Recombinant plasmidpSecTag-opg was successfully transfected into rat MSC cell line by lipofectamine and OPG-expression cell line of rat MSC was selected and confirmed.Conclusion: OPG-expression rat MSC cell line was successfully established and identified.更多还原