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

被引:24
作者
Buo, Atum M. [1 ]
Williams, Mark S. [2 ]
Kerr, Jaclyn P. [1 ]
Stains, Joseph P. [1 ]
机构
[1] Univ Maryland, Sch Med, Dept Orthopaed, Baltimore, MD 21201 USA
[2] Univ Maryland, Sch Med, Dept Microbiol & Immunol, Baltimore, MD 21201 USA
来源
BONE RESEARCH | 2016年 / 4卷
基金
美国国家卫生研究院;
关键词
MEDIATED GENE-TRANSFER; IN-VITRO; CONNEXIN43; VIVO; EXPRESSION; EFFICIENCY; COMPLEXES; REQUIRES; INCREASE; DELETION;
D O I
10.1038/boneres.2016.21
中图分类号
Q813 [细胞工程];
学科分类号
100113 [医学细胞生物学];
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.
引用
收藏
页数:10
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