Effect of theaflavins on the differentiation of rabbit bone marrow mesenchymal stem cells into adipocytes
- VernacularTitle:茶黄素对兔骨髓基质干细胞向脂肪细胞诱导分化的影响
- Author:
Fu LI
;
Xifu SHANG
;
Tao XIE
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2007;0(16):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: Theaflavins are the major constituent of black tea, and have a lot of pharmacological activities, for example, regulating blood fat, anti-oxygen, anti-tumor, resisting cerebrovascular disease, and so on. Previously, theaflavins' effect on human fat metabolism mostly belongs to macro-analysis, but this experiment try to find out the effect of theaflavins on the differentiation of rabbit mesenchymal stem cells (MSCs) into adipocytes in terms of cells. METHODS: The experiment was completed in the center laboratory of Anhui Provincial Hospital Affiliated to Anhui Medical University from May to September in 2007.①Ten rabbits of cleaning grade and 4-8 weeks old were provided by the animal experiment center of Anhui Medical University. The disposal of animals in process of experiment referred to the ethical standard of animals.②The heparinized rabbits were sacrificed in drugged state, then bilateral femur, tibia and humerus were obtained and their soft tissues were removed, cutting epiphysis of two sides including epiphyseal plate. The MSCs were isolated and cultured with whole bone marrow culture method, inoculated at a density of 2?108 L-1. When the cells grew to the fusion of 80%-90%, digesting and passage culture were performed. Cells of the third generation were collected and inoculated at a density of 1?105/cm2 to be cultured for 2 weeks with DMEM adipocyte-induced liquid, which contained recombinant human insulin 10 mg/L, dexamethasone 10-6 mol/L, and IBMX 0.5 mmol/L. Three groups were set up: adipocytes control group in 1 mL adipocyte-induced liquid; theaflavins group with the 500 ?g/L theaflavins in 1 mL adipocyte-induced liquid; blank group in equal amount of ordinary DMEM culture medium.③The cultured cells of the second generation were selected to draw growth curve; oil O staining was used to identify the induced adipocytes and calculate the differentiation efficiency of adipocytes. RESULTS: ①Cell growth curve: MSCs had vigorous reproductive activity, 1 day of the culture was the cell adaptive phase, 3 days was the increased logarithmic phase, and 8 days was the platform phase. After 8 days, cell proliferation quickly stepped down and cells reduced.②Identification of adipocytes by oil O stain: Lipid droplets in adipocytes were stained into salmon pink, and the nucleus was stained into blue. Most of MSCs in adipocyte control group were induced into adipocytes, the differentiation efficiency was (64.8?4.8)%, while that in theaflavins group was only (32.0?3.4)%. No adipocytes obviously formed in blank control group. CONCLUSION: Theaflavins can obviously inhibit differentiation from rabbit MSCs to adipocytes.
