Retroviral-mediated overexpression of human bone morphogenetic protein 2 affects human mesenchymal stem cells during monolayer proliferation: A cautionary note

Background: Retroviral vectors are commonly used for gene transfer applications and they represent an effective way to provide a sustained delivery of a bioactive factor in basic research and tissue engineering applications. Cells that have been transduced with retroviral vectors ex vivo are usually...

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Autores principales:	Neumann,Alexander J, Alini,Mauro, Archer,Charles W, Stoddart,Martin J
Lenguaje:	English
Publicado:	Pontificia Universidad Católica de Valparaíso 2013
Materias:	constitutive expression controls gene transfer monolayer expansion retrovirus
Acceso en línea:	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582013000200007
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spelling	oai:scielo:S0717-345820130002000072013-03-27Retroviral-mediated overexpression of human bone morphogenetic protein 2 affects human mesenchymal stem cells during monolayer proliferation: A cautionary noteNeumann,Alexander JAlini,MauroArcher,Charles WStoddart,Martin J constitutive expression controls gene transfer monolayer expansion retrovirus Background: Retroviral vectors are commonly used for gene transfer applications and they represent an effective way to provide a sustained delivery of a bioactive factor in basic research and tissue engineering applications. Cells that have been transduced with retroviral vectors ex vivo are usually amplified on tissue culture plastic, for a prolonged period of time, in order to obtain sufficient cell numbers prior to the experiment of interest. However, the effect of the transgene product on the transduced cells, during this period of time, is rarely, if ever, investigated. The current study investigated if transduction with a VSG.G pseudotyped retroviral vector expressing human bone morphogenetic protein 2 (Rv.BMP-2) influences the gene expression profile of human bone marrow-derived mesenchymal stem cells (hMSCs) during monolayer proliferation. hMSCs that have been transduced with a VSG.G pseudotyped retroviral vector expressing enhanced green fluorescent protein (Rv.eGFP) served as controls. Results: It was confirmed that Rv.BMP-2 transduced hMSCs produce detectable amounts of bone morphogenetic protein 2 (BMP-2). Gene expression analysis revealed that the hypertrophic marker collagen X was down-regulated by approximately 50% and the chondrogenic marker Aggrecan was elevated almost 9-fold in Rv.BMP-2 transduced hMSCs if compared to Rv.eGFP transduced control cells. Interestingly, the same changes in gene expression were detected when hMSCs were exposed to 100 ng/ml of recombinant human BMP-2 and their gene expression profile was compared to control hMSC. Again, collagen X message was down-regulated and Aggrecan message was up-regulated. Conclusion: These results indicate that, when using integrating vectors and then expanding the cells after transduction, controls need to be carefully planned to ensure the results obtained during the 3D experiments are not due to artefacts created in response to the different cell proliferation conditions employed.info:eu-repo/semantics/openAccessPontificia Universidad Católica de ValparaísoElectronic Journal of Biotechnology v.16 n.2 20132013-03-01text/htmlhttp://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582013000200007en10.2225/vol16-issue2-fulltext-2
institution	Scielo Chile
collection	Scielo Chile
language	English
topic	constitutive expression controls gene transfer monolayer expansion retrovirus
spellingShingle	constitutive expression controls gene transfer monolayer expansion retrovirus Neumann,Alexander J Alini,Mauro Archer,Charles W Stoddart,Martin J Retroviral-mediated overexpression of human bone morphogenetic protein 2 affects human mesenchymal stem cells during monolayer proliferation: A cautionary note
description	Background: Retroviral vectors are commonly used for gene transfer applications and they represent an effective way to provide a sustained delivery of a bioactive factor in basic research and tissue engineering applications. Cells that have been transduced with retroviral vectors ex vivo are usually amplified on tissue culture plastic, for a prolonged period of time, in order to obtain sufficient cell numbers prior to the experiment of interest. However, the effect of the transgene product on the transduced cells, during this period of time, is rarely, if ever, investigated. The current study investigated if transduction with a VSG.G pseudotyped retroviral vector expressing human bone morphogenetic protein 2 (Rv.BMP-2) influences the gene expression profile of human bone marrow-derived mesenchymal stem cells (hMSCs) during monolayer proliferation. hMSCs that have been transduced with a VSG.G pseudotyped retroviral vector expressing enhanced green fluorescent protein (Rv.eGFP) served as controls. Results: It was confirmed that Rv.BMP-2 transduced hMSCs produce detectable amounts of bone morphogenetic protein 2 (BMP-2). Gene expression analysis revealed that the hypertrophic marker collagen X was down-regulated by approximately 50% and the chondrogenic marker Aggrecan was elevated almost 9-fold in Rv.BMP-2 transduced hMSCs if compared to Rv.eGFP transduced control cells. Interestingly, the same changes in gene expression were detected when hMSCs were exposed to 100 ng/ml of recombinant human BMP-2 and their gene expression profile was compared to control hMSC. Again, collagen X message was down-regulated and Aggrecan message was up-regulated. Conclusion: These results indicate that, when using integrating vectors and then expanding the cells after transduction, controls need to be carefully planned to ensure the results obtained during the 3D experiments are not due to artefacts created in response to the different cell proliferation conditions employed.
author	Neumann,Alexander J Alini,Mauro Archer,Charles W Stoddart,Martin J
author_facet	Neumann,Alexander J Alini,Mauro Archer,Charles W Stoddart,Martin J
author_sort	Neumann,Alexander J
title	Retroviral-mediated overexpression of human bone morphogenetic protein 2 affects human mesenchymal stem cells during monolayer proliferation: A cautionary note
title_short	Retroviral-mediated overexpression of human bone morphogenetic protein 2 affects human mesenchymal stem cells during monolayer proliferation: A cautionary note
title_full	Retroviral-mediated overexpression of human bone morphogenetic protein 2 affects human mesenchymal stem cells during monolayer proliferation: A cautionary note
title_fullStr	Retroviral-mediated overexpression of human bone morphogenetic protein 2 affects human mesenchymal stem cells during monolayer proliferation: A cautionary note
title_full_unstemmed	Retroviral-mediated overexpression of human bone morphogenetic protein 2 affects human mesenchymal stem cells during monolayer proliferation: A cautionary note
title_sort	retroviral-mediated overexpression of human bone morphogenetic protein 2 affects human mesenchymal stem cells during monolayer proliferation: a cautionary note
publisher	Pontificia Universidad Católica de Valparaíso
publishDate	2013
url	http://www.scielo.cl/scielo.php?script=sci_arttext&pid=S0717-34582013000200007
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Retroviral-mediated overexpression of human bone morphogenetic protein 2 affects human mesenchymal stem cells during monolayer proliferation: A cautionary note

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