title: Efficient and sustained IGF-1 expression in the adipose tissue-derived stem cells mediated via a lentiviral vector.
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.
authors.affiliation: Department of Cardiology, First Affiliated Hospital of Chinese People's Liberation Army General Hospital, Beijing, 100048, People's Republic of China.
authors.forename: Ting
authors.initials: T
authors.lastname: Chen
authors.forename: Dangsheng
authors.initials: D
authors.lastname: Huang
authors.forename: Guanghui
authors.initials: G
authors.lastname: Chen
authors.forename: Tingshu
authors.initials: T
authors.lastname: Yang
authors.forename: Jun
authors.initials: J
authors.lastname: Yi
authors.forename: Miao
authors.initials: M
authors.lastname: Tian
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D017209:Apoptosis / Q000235:genetics; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D015870:Gene Expression; D023061:Gene Order; D005822:Genetic Vectors / Q000235:genetics*; D006801:Humans; D016130:Immunophenotyping; D007334:Insulin-Like Growth Factor I / Q000235:genetics*; D016086:Lentivirus / Q000235:genetics*; D010641:Phenotype; D051381:Rats; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*; D014161:Transduction, Genetic
chemical_list: D007334:Insulin-Like Growth Factor I