Effect of serum concentrations on differentiation of adipose-derived stem cells into adipocytes / 中国组织工程研究

ABSTRACT

BACKGROUND: Serum concentrations in culture medium influence the proliferation and differentiation of stem cells. OBJECTIVE: To study the proliferation and adipogenic differentiation potential of adipose-derived stem cells under culture with different concentrations of fetal bovine serum. METHODS: The mouse adipose-derived stem cells were harvested from subcutaneous adipose tissue of C57 mice at 7-10 days of age. Concentrations of fetal bovine serum in the culture medium were set to 5%, 10%, 15% and 20%. The passage 3 cells were used for subsequent studies. The proliferation of cells cultured with different serum concentrations was tested using the cell counting kit-8 assay. The adipogenic differentiation potential of cells was evaluated with Bodipy staining. Quantitative RT-PCR and western blot assay were used to determine expression levels of Cebpa and Pparg mRNA as well as C/EBP-α and PPAR-γ protein levels. RESULTS AND CONCLUSION: There was no statistical difference in the cell proliferation among cells cultured with different serum concentrations, even though adipose-derived stem cells cultured with 20% fetal bovine serum showed lower absorbance value at 450 nm than those cultured with 5%, 10% and 15% fetal bovine serum. The expression levels of Cebpa and Pparg mRNA and C/EBP-α and PPAR-γ protein in the 5% and 10% groups were considerably higher than those of the 15% and 20% groups, and the expression levels were highest in the 10% group. To conclude, different concentrations of fetal bovine serum have no significant effects on the proliferation of adipose-derived stem cells, while the adipogenic differentiation potentials of cells are different when cultured with different serum concentrations. High serum concentration can inhibit the adipogenic differentiation of adipose-derived stem cells.