Differentiation of rat bone marrow mesenchymal stem cells to chondrocytes transfected by SOX-9 and insulin-like growth factor-1
10.3969/j.issn.1673-8225.2009.28.014
- VernacularTitle:SOX-9和胰岛素样生长因子1共同转染大鼠骨髓间充质干细胞向软骨细胞的分化
- Author:
Guangbin WANG
;
Hongliang CHEN
;
Dapeng WANG
;
Qin FU
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2009;13(28):5465-5469
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: Although single gene can induce bone marrow mesenchymal stem cells (BMSCs) into chondrocyte, it still has limit action, polygene interaction accord with real demand of body internal environment and is helpful to play synergistic effect and to improve the differentiated effect.OBJECTIVE: To investigate the secretion of target gene and differentiation of BMSCs transfected by BOX-9 and insulin-like growth factor-1 (IGF-1) gene alone and together into chondrocytes.DESIGN, TIME AND SETTING: Two-specimen observation was conducted in the Cell Biological Laboratory of China Medical University between April 2008 and February 2009. MATERIALS: Two 6-week-old healthy male Wistar rats were used for the BMSCs extraction. METHODS: The plasmids pcDNA3.1-1GF-1 and pcDNA3.1- BOX-9 were amplified and extracted. BMSCs of Wistar rats were separated and purified. According to the transfect situation, the BMSCs were divided into 5 groups. ①Non-transfected group:serum-free and double-antibody-free L-DMEM; ②empty vector transfection group: serum-free and double-antibody-free L-DMEM + pcDNA3.1 empty vector and liposome; ③BOX-9 transfection group: serum-free and double-antibody-free L-DMEM +pcDNA3.1-SOX-9 plasmid and liposome; ④IGF-1 transfection group: serum-free and double-antibody-free L-DMEM+pcDNA3.1-1GF-1 plasmid and liposome; ⑤SOX-9 and IGF-1 transfection group: serum-free and double-antibody-free L-DMEM+ pcDNA3.1-1GF-1 plasmid and liposome, pcDNA3.1-SOX-9 plasmid and liposome.MAIN OUTCOME MEASURES: After being transfected, the cells were selected, the proliferation activity was tested by MTT and expression levels of target gene and protein were tested by RT-PCR and Western blot. RESULTS: The proliferation activity (A value) of BMSCs transfected by SOX-9 alone, iGF-1 alone, SOX-9 and IGF-1 together was significantly higher than that of BMSCs in the non-transfected group and empty vector trensfection group (P < 0.01). RT-PCR and Western blot result showed that SOX-9 expression was the highest in the group transfected by SOX-9, the expression of IGF-1 was the highest in the group transfected both by SOX-9 and IGF-1, and the expression of collagen Ⅱ was the highest in the group transfected both by SOX-9 and IGF-1.CONCLUSION: The double gene transfection is supedor to single transfection with regard to the differentiation of BMSCs to chondrocytes and the secretion of extracallular matdx. It is indirectly suggested that IGF-1 induces BMSCs to differentiate into chondrocytes, but the capacity is less than SOX-9.
