Differentiation of rabbit bone marrow mesenchymal stem cells into adipocytes in media containing dexamethasone, 3-isobutyl-1-methylxanthine, insulin and indomethacin
- VernacularTitle:地塞米松、3-异丁基-1-甲基黄嘌呤、胰岛素、吲哚美辛定向诱导兔骨髓间充质干细胞分化为脂肪细胞的研究
- Author:
Yana LI
;
Yan SUN
;
Ling ZHANG
;
Xu LIU
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2008;12(21):4193-4196
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: Adipocytes are derived from preadipocytes, which are induced by mesenchymal stem cell differentiation. This study explored the mechanism of mesenchymal stem cell differentiation into adipocytes and provided a new thinking for preventing and treating obesity.OBJECTIVE: To investigate the isolation and culture of BMSCs from rabbits in vitro, and the method of differentiating into adipocytes.DESIGN: An observational controlled in vitro experiment.SETTING: Department of Histology and Embryology, Binzhou Medical College, and Yunnan Pharmacological Laboratories of Natural Products.MATERIALS: Totally 12 healthy Japanese rabbits aged 6-8 weeks of 200 to 300g of either sex were provided by Experimental Animal Center of Kunming Medical College. Dexamethasone, 3-isobutyl-1-methylxanthine and insulin (Sigma, USA) and indomethacin (ALEXIS, USA) were used in this study.METHODS: Experiments were performed at the Yunnan Pharmacological Laboratories of Natural Products between July 2004 and January 2005. BMSCs were harvested from the femurs, tibias and humerus bones of the rabbits, and then purified and proliferated. After primary culture, the subcultured cells were cultured in media including 1μmol/L dexamethasone, 0.5mmol/L 3-isobutyl-1-methylxanthine, 10mg/L insulin and 0.2mmol/L indomethacin. The disposal was accorded with the animal ethical standards.MAIN OUTCOME MEASURES: Growth curve of the third passage of cells was evaluated by modified MTT method. Salmon pink lipid drop appeared after Oil Red O staining under a light microscope.RESULTS: After 72 hours of adipocyte induction, lipid droplet appeared in BMSCs. With the prolongation of induction time, lipid vacuoles were found and cells changed from fusiform to round or polygon. At week 3, approximately 80% BMSCs were induced into adipocytes as detected by Oil Red O.CONCLUSION: Rabbit BMSCs isolated and cultured in vitro can be induced into adipocytes in the media containing 1μmol/L dexamethasone, 0.5mmol/L 3-isobutyl-1-methylxanthine, 10mg/L insulin and 0.2mmol/L indomethacin.
