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Efficient and sustained IGF-1 expression in the adipose tissue-derived stem cells mediated via a lentiviral vector

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Abstract

The adipose tissue-derived stem cells (ADSCs) represent a significant area of the cell therapy. Genetic modification of ADSCs may further improve their therapeutic potential. Here, we aimed to generate a lentiviral vector expressing insulin-like growth factor-I (IGF-1) and investigate the impact of IGF-1 transduction on the properties of cultured ADSCs. Isolated rat ADSCs were assessed by flow cytometric analysis. IGF-1 was cloned and inserted into the pLenO-DCE plasmid to acquire pLenO-DCE-IGF-1 plasmid. Lentivirus was enveloped with pRsv-REV, pMDlg-pRRE and pMD2G plasmids in 293T cells. The ADSCs were transfected with the vectors. And then IGF-1-induced anti-apoptosis was evaluated by annexin V-FITC. Besides, proliferation of cells was detected by MTT assay and EdU. Moreover, Akt phosphorylation was evaluated by Western blotting analysis. Stable expression of IGF-1 in ADSCs was confirmed. ADSCs were positive for CD90 and CD29, but negative for CD31, CD34 and CD45. The transduction of IGF-1 to the ADSCs caused a dramatic increase in P-Akt expression. Over-expression of IGF-1 in ADSCs could improve the paracine of IGF-1 in a time-dependent manner, but could not promote the proliferation of ADSCs. This study indicated that lentiviral vectors offered a promising mean of delivering IGF-1 to the ADSCs. Lentiviral-mediated over-expression of therapeutic IGF-1 gene in ADSCs could prolong the anti-apoptosis effect of IGF-1, which might be induced by the activation of the PI3K/Akt pathway. And our data would improve the efficacy of ADSC-based therapies.

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Acknowledgements

We wish to express our warm thanks to the support from National high-technology research and develop project (863) of China No. 2011AA020101 and all the authors who contributed to the research. National high-technology research and develop project (863) of China No. 2011AA020101.

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We have no conflict of interest to state.

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Authors and Affiliations

  1. Department of Cardiology, First Affiliated Hospital of Chinese People’s Liberation Army General Hospital, Beijing, 100048, People’s Republic of China

    Ting Chen, Dangsheng Huang & Miao Tian

  2. Department of Cardiology, Chinese People’s Liberation Army General Hospital, Fuxin Road No. 28, Haidian District, Beijing, 100853, People’s Republic of China

    Guanghui Chen & Tingshu Yang

  3. Department of Cardiac Laboratory, Chinese People’s Liberation Army General Hospital, Beijing, 100853, People’s Republic of China

    Jun Yi

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  1. Ting Chen
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  2. Dangsheng Huang
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  3. Guanghui Chen
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  4. Tingshu Yang
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  5. Jun Yi
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  6. Miao Tian
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Correspondence to Guanghui Chen.

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Ting Chen and Dangsheng Huang have contributed equally to this work.

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Chen, T., Huang, D., Chen, G. et al. Efficient and sustained IGF-1 expression in the adipose tissue-derived stem cells mediated via a lentiviral vector. J Mol Hist 46, 1–11 (2015). https://doi.org/10.1007/s10735-014-9599-7

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  • DOI: https://doi.org/10.1007/s10735-014-9599-7

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