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

发布时间：2018-07-10 16:23

本文选题：骨保护素 + 基因　；参考：《安徽医科大学》2006年硕士论文

【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。
[Abstract]:Culture of rat bone marrow stromal cell lines modified with osteoprotegerin gene Objective: to culture rat bone marrow stromal cell lines modified by OPG gene and to identify the expression products. It provides material basis for genetic engineering in the treatment of periodontal disease. Methods: bone marrow stromal cells were isolated from adult SD rat bone marrow by density gradient centrifugation. The target gene was transfected into rat bone marrow stromal cells by liposome method using the constructed pSecTag-opg eukaryotic expression shuttle vector. The expression of osteoprotegerin was confirmed by immunohistochemistry and western blot. Results: rat bone marrow stromal cells were isolated and cultured. The target gene was transfected by liposome method and a stable transfected cell line was established. The expression of osteoprotegerin was detected by immunohistochemistry and western blot in rat bone marrow stromal cells transfected with OPG gene. Conclusion: rat bone marrow stromal cell lines secreting OPG were successfully cultured in this experiment.
【学位授予单位】：安徽医科大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R329.2

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