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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...
Autores principales: | , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2016
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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. |
format | Online Article Text |
id | pubmed-4977485 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
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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