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

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 cy...

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Detalles Bibliográficos
Autores principales: Buo, Atum M, Williams, Mark S, Kerr, Jaclyn P, Stains, Joseph P
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4977485/
https://www.ncbi.nlm.nih.gov/pubmed/27547486
http://dx.doi.org/10.1038/boneres.2016.21
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author Buo, Atum M
Williams, Mark S
Kerr, Jaclyn P
Stains, Joseph P
author_facet Buo, Atum M
Williams, Mark S
Kerr, Jaclyn P
Stains, Joseph P
author_sort Buo, Atum M
collection PubMed
description 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.
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spelling pubmed-49774852016-08-19 A cost-effective method to enhance adenoviral transduction of primary murine osteoblasts and bone marrow stromal cells Buo, Atum M Williams, Mark S Kerr, Jaclyn P Stains, Joseph P Bone Res Article 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. Nature Publishing Group 2016-08-09 /pmc/articles/PMC4977485/ /pubmed/27547486 http://dx.doi.org/10.1038/boneres.2016.21 Text en Copyright © 2016 Sichuan University http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Buo, Atum M
Williams, Mark S
Kerr, Jaclyn P
Stains, Joseph P
A cost-effective method to enhance adenoviral transduction of primary murine osteoblasts and bone marrow stromal cells
title A cost-effective method to enhance adenoviral transduction of primary murine osteoblasts and bone marrow stromal cells
title_full A cost-effective method to enhance adenoviral transduction of primary murine osteoblasts and bone marrow stromal cells
title_fullStr A cost-effective method to enhance adenoviral transduction of primary murine osteoblasts and bone marrow stromal cells
title_full_unstemmed A cost-effective method to enhance adenoviral transduction of primary murine osteoblasts and bone marrow stromal cells
title_short A cost-effective method to enhance adenoviral transduction of primary murine osteoblasts and bone marrow stromal cells
title_sort cost-effective method to enhance adenoviral transduction of primary murine osteoblasts and bone marrow stromal cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4977485/
https://www.ncbi.nlm.nih.gov/pubmed/27547486
http://dx.doi.org/10.1038/boneres.2016.21
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