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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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Detalles Bibliográficos
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
Descripción
Sumario: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.