The Effect Of Baicalin On The Differentiation Of D3 Mouse Embryonic Stem Cell Into Cardiomyocytes-like Cells

Objective: To evaluate the effect of baicalin on the differentiation of D3 Mouse Embryonic stem cell into cardiomyocytes-like cells in vitro.Method: We cultured ES cell in 96-well plate for 8 days in vitro without LIF and detected the effect of baicalin with different concentrations on the proliferation of ES cell, then we selected the suitable concentrations for differentiation. With the method of "hanging drops-suspension culture-adherence culture", baicalin at 3 concentrations was used to induce the differentiation of ESC. We observed the morphology of cells with invert microscope and counted the beating EBs everyday since the EB adhered to the plasitic plates, we detected the expression of sarcomere-specificα-actinin with immunocytochemistry and investigated the expression ofα-MHC on RNA level with RT-PCR. Finally, we detected the cardiac differentiation rate with flow cytometry.Result: MTT showed that baicalin at concentration of 1μmol/L, 10μmol/L, 102μmol/L,103μmol/L obviously inhibited the proliferation of mES-D3 cells and 10-2μmol/L had the lowest effect on the proliferation of mES-D3 cells. By observing the morphology of EB, We found that compared with the control, the EBs of the groups treated with baicalin were much fewer and smaller. However, flow cytometry showed that the 1μmol/L group has the highest cardiac differentiation rate. Furthermore the immunocytochemistry and RT-PCR confirmed the expression ofα-actinin andα-MHC at protein level and mRNA level respectively.Conclusion: Baicalin at High concentration could obviousely inhibit the proliferation of mES-D3 cells, and baicalin at 1μmol/L may improve the cardiac differentiation rate, the differentiation rate was 21.45%.