Effects of SP600125 at various concentrations on proliferation and osteogenesis of human adipose-derived stem cells in vitro

Abstract

Objective To elucidate the effects of SP600125 at different concentrations on the proliferation and osteo-differentiation of human adipose-derived stem cells (hASCs). Methods The hASCs harvested were cocultured with SP600125 at concentrations of 0 μmol/L, 1 μmol/L, 5 μmol/L and 10 μmol/L in growth medium (OM group) and in osteogenesis medium (OM group), respectively. The DNA quantitative assay was carried out to evaluate proliferation of the hASCs; flow cytometry was used to determine the effect of SP600125 on the cell cycles of hASCs; Alkaline phosphatase level (ALP) and calcium deposition tests were conducted to observe the effects of SP600125 at different concentrations on osteogenic differentiation of the hASCs. Results The proliferation of hASCs was inhibited by 42.1% when the cells were cocultured with SP600125 at the concentration of 10 μmol/L; the suppression decreased with decreased concentration of SP600125. The hASCs of phase G0/G1 in GM cocultured with SP600125 at the concentration of 10 μmol/L were more than those in GM cocultured with dimethylsulfoxide at the same concentration. ALP test revealed that after 10 days of culture in vitro the staining was more and more weakened and scattered and the ALP activity was more and more decreased with the increased concentration of SP600125. The extracellular calcium deposition of hASCs after 14 days of culture in vitro showed that the size and number of calcium nodules decreased with the increased concentration of SP600125. Conclusion SP600125 can suppress the proliferation and osteogenic differentiation of hASCs in vitro. Key words: Human; Adipocytes; Cell proliferation; Osteogenic differentiation; SP600125