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

J Mol Histol. 2015 Feb;46(1):1-11. doi: 10.1007/s10735-014-9599-7. Epub 2014 Dec 3.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adipose Tissue / cytology*
  • Animals
  • Apoptosis / genetics
  • Cell Differentiation
  • Cell Line, Tumor
  • Cell Proliferation
  • Gene Expression*
  • Gene Order
  • Genetic Vectors / genetics*
  • Humans
  • Immunophenotyping
  • Insulin-Like Growth Factor I / genetics*
  • Lentivirus / genetics*
  • Phenotype
  • Rats
  • Stem Cells / cytology*
  • Stem Cells / metabolism*
  • Transduction, Genetic

Substances

  • Insulin-Like Growth Factor I