Effect of 17β-estradiol on Ca2+ channels during osteogenic differentiation of rat bone marrow mesenchymal stem cells / 吉林大学学报(医学版)

ABSTRACT

Objective: To observe the effect of 17β-estradiol (17β-E2) on the calcium (Ca2) channels during the osteogenic differentiation of rat bone marrow mesenchymal stem cells (MSCs), and to elucidate the mechanism of 17β-E2 in the osteogenic differentiation of MSCs. Methods: The MSCs were separated by density gradient centrifugation and adherent screening, and passaged for 3 times continuously to induce osteoblast differentiation. The MSCs were divided into control group [cultivated in osteoblast culture medium alone (OBM)] and different doses of 17β-E2 groups (added with 0. 1, 1.0, 10. 0, and 100. 0 pmol · L-1 17β-E2 in OBM, respectively). On the 14th day of osteogenic induction, the cells in each group were stained with Fluo-3/AM, and the Ca2 levels were determinated by laser scanning confocal microscope; the mean fluorescence intensity (MFD was used to respresent the level of Ca2. Whole-cell Ca2 currents were recorded using whole-cell patch clamp technique under different conditions. Results: The MSCs with fibroblast-like cells, oval nuclei and visible nucleoli were successfully isolated by density gradient centrifugation and adherent screening. The subcultured MSCs grew vigorously and maintained the morphological characteristics of primary cells. Following the increase of 17β-E2 concentration, the Fluo-3 fluorescence staining intensity of Ca2 in each group was also gradually increased, especially in 100. 0 pmol · L-1 17β-E2 group. Compared with control group, the MFI of Ca' and the current peak values of Ca' in 10. 0 and 100. 0 pmol · L-1 17β-E2 groups were increased (P0. 05). Conclusion: The cells isolated by density gradient and adherent screening method are the rat MScs. 17β-E2 plays a role in promoting osteogenesis by enhancing the opening of Ca' channels in the MSCs and the inward current of calcium ions in a dose-dependent manner.