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Adenovirus-Mediated Gene Transfer in Mesenchymal Stem Cells Can Be Significantly Enhanced by the Cationic Polymer Polybrene
Mesenchymal stem cells (MSCs) are multipotent progenitors, which can undergo self-renewal and give rise to multi-lineages. A great deal of attentions have been paid to their potential use in regenerative medicine as potential therapeutic genes can be introduced into MSCs. Genetic manipulations in MS...
Autores principales: | , , , , , , , , , , , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2014
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3962475/ https://www.ncbi.nlm.nih.gov/pubmed/24658746 http://dx.doi.org/10.1371/journal.pone.0092908 |
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author | Zhao, Chen Wu, Ningning Deng, Fang Zhang, Hongmei Wang, Ning Zhang, Wenwen Chen, Xian Wen, Sheng Zhang, Junhui Yin, Liangjun Liao, Zhan Zhang, Zhonglin Zhang, Qian Yan, Zhengjian Liu, Wei Wu, Di Ye, Jixing Deng, Youlin Zhou, Guolin Luu, Hue H. Haydon, Rex C. Si, Weike He, Tong-Chuan |
author_facet | Zhao, Chen Wu, Ningning Deng, Fang Zhang, Hongmei Wang, Ning Zhang, Wenwen Chen, Xian Wen, Sheng Zhang, Junhui Yin, Liangjun Liao, Zhan Zhang, Zhonglin Zhang, Qian Yan, Zhengjian Liu, Wei Wu, Di Ye, Jixing Deng, Youlin Zhou, Guolin Luu, Hue H. Haydon, Rex C. Si, Weike He, Tong-Chuan |
author_sort | Zhao, Chen |
collection | PubMed |
description | Mesenchymal stem cells (MSCs) are multipotent progenitors, which can undergo self-renewal and give rise to multi-lineages. A great deal of attentions have been paid to their potential use in regenerative medicine as potential therapeutic genes can be introduced into MSCs. Genetic manipulations in MSCs requires effective gene deliveries. Recombinant adenoviruses are widely used gene transfer vectors. We have found that although MSCs can be infected in vitro by adenoviruses, high virus titers are needed to achieve high efficiency. Here, we investigate if the commonly-used cationic polymer Polybrene can potentiate adenovirus-mediated transgene delivery into MSCs, such as C2C12 cells and iMEFs. Using the AdRFP adenovirus, we find that AdRFP transduction efficiency is significantly increased by Polybrene in a dose-dependent fashion peaking at 8 μg/ml in C2C12 and iMEFs cells. Quantitative luciferase assay reveals that Polybrene significantly enhances AdFLuc-mediated luciferase activity in C2C12 and iMEFs at as low as 4 μg/ml and 2 μg/ml, respectively. FACS analysis indicates that Polybrene (at 4 μg/ml) increases the percentage of RFP-positive cells by approximately 430 folds in AdRFP-transduced iMEFs, suggesting Polybrene may increase adenovirus infection efficiency. Furthermore, Polybrene can enhance AdBMP9-induced osteogenic differentiation of MSCs as early osteogenic marker alkaline phosphatase activity can be increased more than 73 folds by Polybrene (4 μg/ml) in AdBMP9-transduced iMEFs. No cytotoxicity was observed in C2C12 and iMEFs at Polybrene up to 40 μg/ml, which is about 10-fold higher than the effective concentration required to enhance adenovirus transduction in MSCs. Taken together, our results demonstrate that Polybrene should be routinely used as a safe, effective and inexpensive augmenting agent for adenovirus-mediated gene transfer in MSCs, as well as other types of mammalian cells. |
format | Online Article Text |
id | pubmed-3962475 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2014 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-39624752014-03-24 Adenovirus-Mediated Gene Transfer in Mesenchymal Stem Cells Can Be Significantly Enhanced by the Cationic Polymer Polybrene Zhao, Chen Wu, Ningning Deng, Fang Zhang, Hongmei Wang, Ning Zhang, Wenwen Chen, Xian Wen, Sheng Zhang, Junhui Yin, Liangjun Liao, Zhan Zhang, Zhonglin Zhang, Qian Yan, Zhengjian Liu, Wei Wu, Di Ye, Jixing Deng, Youlin Zhou, Guolin Luu, Hue H. Haydon, Rex C. Si, Weike He, Tong-Chuan PLoS One Research Article Mesenchymal stem cells (MSCs) are multipotent progenitors, which can undergo self-renewal and give rise to multi-lineages. A great deal of attentions have been paid to their potential use in regenerative medicine as potential therapeutic genes can be introduced into MSCs. Genetic manipulations in MSCs requires effective gene deliveries. Recombinant adenoviruses are widely used gene transfer vectors. We have found that although MSCs can be infected in vitro by adenoviruses, high virus titers are needed to achieve high efficiency. Here, we investigate if the commonly-used cationic polymer Polybrene can potentiate adenovirus-mediated transgene delivery into MSCs, such as C2C12 cells and iMEFs. Using the AdRFP adenovirus, we find that AdRFP transduction efficiency is significantly increased by Polybrene in a dose-dependent fashion peaking at 8 μg/ml in C2C12 and iMEFs cells. Quantitative luciferase assay reveals that Polybrene significantly enhances AdFLuc-mediated luciferase activity in C2C12 and iMEFs at as low as 4 μg/ml and 2 μg/ml, respectively. FACS analysis indicates that Polybrene (at 4 μg/ml) increases the percentage of RFP-positive cells by approximately 430 folds in AdRFP-transduced iMEFs, suggesting Polybrene may increase adenovirus infection efficiency. Furthermore, Polybrene can enhance AdBMP9-induced osteogenic differentiation of MSCs as early osteogenic marker alkaline phosphatase activity can be increased more than 73 folds by Polybrene (4 μg/ml) in AdBMP9-transduced iMEFs. No cytotoxicity was observed in C2C12 and iMEFs at Polybrene up to 40 μg/ml, which is about 10-fold higher than the effective concentration required to enhance adenovirus transduction in MSCs. Taken together, our results demonstrate that Polybrene should be routinely used as a safe, effective and inexpensive augmenting agent for adenovirus-mediated gene transfer in MSCs, as well as other types of mammalian cells. Public Library of Science 2014-03-21 /pmc/articles/PMC3962475/ /pubmed/24658746 http://dx.doi.org/10.1371/journal.pone.0092908 Text en © 2014 Zhao et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Zhao, Chen Wu, Ningning Deng, Fang Zhang, Hongmei Wang, Ning Zhang, Wenwen Chen, Xian Wen, Sheng Zhang, Junhui Yin, Liangjun Liao, Zhan Zhang, Zhonglin Zhang, Qian Yan, Zhengjian Liu, Wei Wu, Di Ye, Jixing Deng, Youlin Zhou, Guolin Luu, Hue H. Haydon, Rex C. Si, Weike He, Tong-Chuan Adenovirus-Mediated Gene Transfer in Mesenchymal Stem Cells Can Be Significantly Enhanced by the Cationic Polymer Polybrene |
title | Adenovirus-Mediated Gene Transfer in Mesenchymal Stem Cells Can Be Significantly Enhanced by the Cationic Polymer Polybrene |
title_full | Adenovirus-Mediated Gene Transfer in Mesenchymal Stem Cells Can Be Significantly Enhanced by the Cationic Polymer Polybrene |
title_fullStr | Adenovirus-Mediated Gene Transfer in Mesenchymal Stem Cells Can Be Significantly Enhanced by the Cationic Polymer Polybrene |
title_full_unstemmed | Adenovirus-Mediated Gene Transfer in Mesenchymal Stem Cells Can Be Significantly Enhanced by the Cationic Polymer Polybrene |
title_short | Adenovirus-Mediated Gene Transfer in Mesenchymal Stem Cells Can Be Significantly Enhanced by the Cationic Polymer Polybrene |
title_sort | adenovirus-mediated gene transfer in mesenchymal stem cells can be significantly enhanced by the cationic polymer polybrene |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3962475/ https://www.ncbi.nlm.nih.gov/pubmed/24658746 http://dx.doi.org/10.1371/journal.pone.0092908 |
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