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Lentiviral-mediated ephrin B2 gene modification of rat bone marrow mesenchymal stem cells

OBJECTIVE: To determine the effect of the upregulation or knockdown of the ephrinB2 (Efnb2) gene and the effect of EphB4/EphrinB2 signalling in rat bone marrow mesenchymal stem cells (BMSCs). METHODS: Rat BMSCs were infected with lentivirus vectors carrying EphrinB2 and shRNA-EphrinB2. EphrinB2 mRNA...

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Autores principales: Zhu, Min, Hua, Yu, Tang, Jian, Zhao, Xiaoke, Zhang, Ling, Zhang, Yue
Formato: Online Artículo Texto
Lenguaje:English
Publicado: SAGE Publications 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6683898/
https://www.ncbi.nlm.nih.gov/pubmed/31122164
http://dx.doi.org/10.1177/0300060519843023
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author Zhu, Min
Hua, Yu
Tang, Jian
Zhao, Xiaoke
Zhang, Ling
Zhang, Yue
author_facet Zhu, Min
Hua, Yu
Tang, Jian
Zhao, Xiaoke
Zhang, Ling
Zhang, Yue
author_sort Zhu, Min
collection PubMed
description OBJECTIVE: To determine the effect of the upregulation or knockdown of the ephrinB2 (Efnb2) gene and the effect of EphB4/EphrinB2 signalling in rat bone marrow mesenchymal stem cells (BMSCs). METHODS: Rat BMSCs were infected with lentivirus vectors carrying EphrinB2 and shRNA-EphrinB2. EphrinB2 mRNA and protein levels were quantified. At 28 days of culture with neuronal cell-conditioned differentiation medium, levels of microtubule-associated protein 2 (MAP2), CD133 and nestin were detected in EphrinB2/BMSCs and shEphrinB2/BMSCs using quantitative polymerase chain reaction and immunofluorescence. The ability of these cells to migrate was evaluated using a transwell assay. RESULTS: BMSCs were successfully isolated as indicated by their CD90+ CD29+ CD34– CD45– phenotype. Three days after ephrinB2 transduction, BMSC cell bodies began to shrink and differentiate into neuron-like cells. At 28 days, levels of MAP2, CD133 and nestin, as well as the number of migratory cells, were higher in lenti-EphrinB2-BMSCs than in the two control groups. The shEphrinB2/BMSCs had reduced levels of MAP2, CD133 and nestin; and a lower rate of cell migration. Similarly, increased levels of Grb4 andp21-activated kinase in the EphB4/EphrinB2 reverse signalling pathway were observed by Western blot. CONCLUSIONS: LV-EphrinB2 can be efficiently transduced into BMSCs, which then differentiate into neuron-like cells.
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spelling pubmed-66838982019-08-19 Lentiviral-mediated ephrin B2 gene modification of rat bone marrow mesenchymal stem cells Zhu, Min Hua, Yu Tang, Jian Zhao, Xiaoke Zhang, Ling Zhang, Yue J Int Med Res Pre-Clinical Research Reports OBJECTIVE: To determine the effect of the upregulation or knockdown of the ephrinB2 (Efnb2) gene and the effect of EphB4/EphrinB2 signalling in rat bone marrow mesenchymal stem cells (BMSCs). METHODS: Rat BMSCs were infected with lentivirus vectors carrying EphrinB2 and shRNA-EphrinB2. EphrinB2 mRNA and protein levels were quantified. At 28 days of culture with neuronal cell-conditioned differentiation medium, levels of microtubule-associated protein 2 (MAP2), CD133 and nestin were detected in EphrinB2/BMSCs and shEphrinB2/BMSCs using quantitative polymerase chain reaction and immunofluorescence. The ability of these cells to migrate was evaluated using a transwell assay. RESULTS: BMSCs were successfully isolated as indicated by their CD90+ CD29+ CD34– CD45– phenotype. Three days after ephrinB2 transduction, BMSC cell bodies began to shrink and differentiate into neuron-like cells. At 28 days, levels of MAP2, CD133 and nestin, as well as the number of migratory cells, were higher in lenti-EphrinB2-BMSCs than in the two control groups. The shEphrinB2/BMSCs had reduced levels of MAP2, CD133 and nestin; and a lower rate of cell migration. Similarly, increased levels of Grb4 andp21-activated kinase in the EphB4/EphrinB2 reverse signalling pathway were observed by Western blot. CONCLUSIONS: LV-EphrinB2 can be efficiently transduced into BMSCs, which then differentiate into neuron-like cells. SAGE Publications 2019-05-24 2019-07 /pmc/articles/PMC6683898/ /pubmed/31122164 http://dx.doi.org/10.1177/0300060519843023 Text en © The Author(s) 2019 http://creativecommons.org/licenses/by-nc/4.0/ Creative Commons Non Commercial CC BY-NC: This article is distributed under the terms of the Creative Commons Attribution-NonCommercial 4.0 License (http://www.creativecommons.org/licenses/by-nc/4.0/) which permits non-commercial use, reproduction and distribution of the work without further permission provided the original work is attributed as specified on the SAGE and Open Access pages (https://us.sagepub.com/en-us/nam/open-access-at-sage).
spellingShingle Pre-Clinical Research Reports
Zhu, Min
Hua, Yu
Tang, Jian
Zhao, Xiaoke
Zhang, Ling
Zhang, Yue
Lentiviral-mediated ephrin B2 gene modification of rat bone marrow mesenchymal stem cells
title Lentiviral-mediated ephrin B2 gene modification of rat bone marrow mesenchymal stem cells
title_full Lentiviral-mediated ephrin B2 gene modification of rat bone marrow mesenchymal stem cells
title_fullStr Lentiviral-mediated ephrin B2 gene modification of rat bone marrow mesenchymal stem cells
title_full_unstemmed Lentiviral-mediated ephrin B2 gene modification of rat bone marrow mesenchymal stem cells
title_short Lentiviral-mediated ephrin B2 gene modification of rat bone marrow mesenchymal stem cells
title_sort lentiviral-mediated ephrin b2 gene modification of rat bone marrow mesenchymal stem cells
topic Pre-Clinical Research Reports
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6683898/
https://www.ncbi.nlm.nih.gov/pubmed/31122164
http://dx.doi.org/10.1177/0300060519843023
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