BACKGROUND: The low survival rate of transplanted bone marrow mesenchymal stem cells (BMSCs) limits their therapeutic action. It is reasonable for us to look for a suitable drug to promote it proliferation.
OBJECTIVE: To investigate whether Dushentang may enhance the proliferation of BMSCs, and to screen the optimal treating concentration.
METHODS: After Wistar rats were anesthetized, BMSCs were obtained from the femoral and tibial bones, and then cultured to the fifth passage. Surface molecule markers of BMSCs were examined after 5 passages by flow cytometry. Cells were seeded in a 96-well microtiter plate at a density of 1×104 cells/well. Cells were then treated with Dushentang at concentration 10, 5, 2.5, 1.25… and 0.019 5 g/L for 1 or 7 days. Proliferation was determined after Dushentang treatment using colorimetry based on the uptake of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide salt (MTT) by viable cells.
RESULTS AND CONCLUSION: ①Most primary BMSCs adhered to the wall at 2 days after culture, which proliferated faster after passaged, and the fifth passage of cells were mostly purified into BMSCs, spread radially or vortex-likely. ②Surface molecule markers of the fifth passage of BMSCs were examined. The isolated cell purities were 86.8% for CD90, 94.7% for CD44, but negative for CD34 (98.7%) and CD45 (97.1%). ③Proliferation of BMSCs might be enhanced by Dushentang treatment with the concentration from 0.625, 0.312 5, 0.156, 0.078, 0.039, and 0.019 5 g/L, and the optimal concentration was 0.078 g/L.