A cost-effective method to enhance adenoviral transduction of primary murine osteoblasts and bone marrow stromal cells

查看全文 作　　者：Atum M [1]Buo;Mark S [2]Williams;Jaclyn P [1]Kerr;Joseph P [1]Stains 机构地区：[1]Department of Orthopaedics, University of Maryland School of Medicine;[2]Department of Microbiology and Immunology, University of Maryland School of Medicine 出　　处：《Bone Research》索引2016年第4卷第2期,共10页 基　　金：supported by grants, R01-AR063631 (JPS) and F31-AR064673 (AMB), from the National Institutes of Health/National Institute for Arthritis, Musculoskeletal and Skin Diseases 摘　　要：We report here a method for the use of poly-L-lysine(PLL) to markedly improve the adenoviral transduction efficiency of primary murine osteoblasts and bone marrow stromal cells(BMSCs) in culture and in situ,which are typically difficult to transduce. We show by fluorescence microscopy and flow cytometry that the addition of PLL to the viral-containing medium significantly increases the number of green fluorescence protein(GFP)-positive osteoblasts and BMSCs transduced with an enhanced GFP-expressing adenovirus. We also demonstrate that PLL can greatly enhance the adenoviral transduction of osteoblasts and osteocytes in situ in ex vivo tibia and calvaria, as well as in long bone fragments. In addition, we validate that PLL can improve routine adenoviral transduction studies by permitting the use of low multiplicities of infection to obtain the desired biologic effect. Ultimately, the use of PLL to facilitate adenoviral gene transfer in osteogenic cells can provide a cost-effective means of performing efficient gene transfer studies in the context of bone research. 关 键 词：骨髓基质细胞 成骨细胞 成本效益 腺病毒 转导 增强型绿色荧光蛋白 小鼠 骨髓间充质干细胞