Figure 1
Estradiol and phytoestrogens enhance osteogenic and adipogenic differentiation of bone marrow-derived mesenchymal stem cells and adipose-derived stromal/stem cells. (A) to (D) Bone marrow-derived mesenchymal stem cells (BMSCs; n = 6) and adipose-derived stromal/stem cells (ASCs; n = 6) were cultured in charcoal dextran-stripped osteogenic differentiation medium (CDS-ODM) or charcoal dextran-stripped adipogenic differentiation medium (CDS-ADM) and simultaneously delivered vehicle (dimethylsulfoxide (DMSO)), 17β-estradiol (E2; 10 nM), daidzein (1 μM), or genistein (1 μM). (A) After 14 days, cells treated with E2, daidzein, or genistein and induced with CDS-ODM were stained with alizarin red. Scale bar represents 100 μm. (B) Stains were eluted with 10% cetylpyridinium chloride (CPC) to quantify the amount of alizarin red staining and measured at 590 nm. Osteogenic differentiation was determined relative to vehicle-treated cells (normalized to 1.0). (C) After 14 days, cells induced with CDS-ADM were stained with oil red O. Scale bar represents 100 μm. (D) To quantify the amount of oil red O staining, cells were eluted with isopropanol and measured at 544 nm. Adipogenic differentiation was determined relative to vehicle-treated cells (normalized to 1.0). (E), (F) BMSCs (n = 6) and ASCs (n = 6) were cultured in CDS-ODM and simultaneously delivered vehicle (DMSO), E2 (10 nM), daidzein (1 μM), daidzein analog 2c (1 μM), daidzein analog 2g (1 μM), or daidzein analog 2l (1 μM). (E) After 14 days, cells induced with CDS-ODM were stained with alizarin red. Scale bar represents 100 μm. (F) To quantify the amount of alizarin red staining in BMSCs and ASCs treated with vehicle, E2, daidzein, or daidzein analogs, cells were eluted with 10% CPC and measured at 590 nm. Osteogenic differentiation was determined relative to vehicle-treated cells normalized to 1.0. Bars, ± standard deviation. *P < 0.05; **P < 0.01 compared with vehicle-treated cells. ###P < 0.001 compared with E2-treated cells. ΨΨΨP < 0.001 compared with daidzein-treated cells.