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Generation and characterization of transgenic mouse mesenchymal stem cell lines expressing hIGF-1 or hG-CSF

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Abstract

Mesenchymal stem cells (MSC) are promising tools in the fields of cell therapy and regenerative medicine. In addition to their differentiation potential, MSC have the ability to secrete bioactive molecules that stimulate tissue regeneration. Thus, the overexpression of cytokines and growth factors may enhance the therapeutic effects of MSC. Here we generated and characterized mouse bone marrow MSC lines overexpressing hG-CSF or hIGF-1. MSC lines overexpressing hG-CSF or hIGF-1 were generated through lentiviral vector mediated gene transfer. The expression of hG-CSF or hIGF-1 genes in the clones produced was quantified by qRT-PCR, and the proteins were detected in the cell supernatants by ELISA. The cell lines displayed cell surface markers and differentiation potential into adipocytes, osteocytes and chondrocytes similar to the control MSC cell lines, indicating the conservation of their phenotype even after genetic modification. IGF-1 and G-CSF transgenic cells maintained immunosuppressive activity. Finally, we performed a comparative gene expression analysis by qRT-PCR array in the cell lines expressing hIGF-1 and hG-CSF when compared to the control cells. Our results demonstrate that the cell lines generated may be useful tools for cell therapy and are suitable for testing in disease models.

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Acknowledgements

This work was supported by The National Council for Scientific and Technological Development (CNPq), The Foundation of Support for Research of the State of Bahia (FAPESB), and Funding Authority for Studies and Projects (FINEP).

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Authors and Affiliations

  1. Center for Biotechnology and Cell Therapy, Hospital São Rafael, Salvador, BA, 41253-190, Brazil

    Gabrielle V. M. Gonçalves, Daniela N. Silva, Rejane H. Carvalho, Bruno S. F. Souza, Kátia Nunes da Silva, Juliana F. Vasconcelos, Bruno D. Paredes, Carolina K. V. Nonaka, Ricardo Ribeiro-dos-Santos & Milena B. P. Soares

  2. Gonçalo Moniz Institute, Oswaldo Cruz Foundation (FIOCRUZ), Rua Waldemar Falcão, 121, Salvador, BA, 40296-710, Brazil

    Daniela N. Silva, Bruno S. F. Souza, Kátia Nunes da Silva, Juliana F. Vasconcelos & Milena B. P. Soares

  3. National Institute of Science and Technology for Regenerative Medicine, Rio de Janeiro, RJ, Brazil

    Bruno S. F. Souza, Bruno D. Paredes, Ricardo Ribeiro-dos-Santos & Milena B. P. Soares

Authors
  1. Gabrielle V. M. Gonçalves
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  2. Daniela N. Silva
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  3. Rejane H. Carvalho
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  4. Bruno S. F. Souza
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  5. Kátia Nunes da Silva
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  6. Juliana F. Vasconcelos
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  7. Bruno D. Paredes
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  8. Carolina K. V. Nonaka
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  9. Ricardo Ribeiro-dos-Santos
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  10. Milena B. P. Soares
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Correspondence to Milena B. P. Soares.

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Gonçalves, G.V.M., Silva, D.N., Carvalho, R.H. et al. Generation and characterization of transgenic mouse mesenchymal stem cell lines expressing hIGF-1 or hG-CSF . Cytotechnology 70, 577–591 (2018). https://doi.org/10.1007/s10616-017-0131-2

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  • DOI: https://doi.org/10.1007/s10616-017-0131-2

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