Differentiation of bone marrow stem cells into the cardiomyocyte-like cells induced by fetal cardiac supernatant with 6-bromoindirubin-3-oxime

ZHOU Cheng-bin; LUI Raphael C; QI Zhou-cuo; YAO Li-ming; ZHUANG Jian

摘要：Background Our previous study showed the 150 mg/mL fetal cardiac supernatant (FCS) could induce differentiation of BMSCs into cardiomyocye-like cells without cardiomyocyte touch, but differentiation efficiency is not high enough.Inhibition of glycogen synthase kinase-3 enhanced the proliferation and survives of stem cells.We tested if 6-bromoindirubin-3-oxime (BIO,glycogen synthase kinase-3 inhibitor) enhances the effects of FCS on differentiation of BMSCs and explore the growth factors in FCS. Methods BMSCs were isolated from the femur and tibia of four-week-old male Sprague-Dawley rats and co-cultured with FCS (150 mg/mL) that was made from fetal hearts from nineteen-day pregnant Wistar rats.BIO with different concentration (0,1,10,and 100 nM) was introduced in culture dishes.Transforming growth factor beta 1 (TGF-β1),bone morphogenetic protein 2 (BMP-2) and Akt in cardiac supernatant and culture medium were assayed with ELISA methods.Results After co-culturing with FCS, beating myotubes were observed in 25.9 ％ BMSCs dishes after 1 to 2 weeks' culture. The levels of TGF-β1 and BMP-2 in FCS concentrations were no more than that in young and adult cardiac supernatant. All BIO groups significantly enhanced the effects of FCS on differentiation of BMSCs into the cardiomyocyte-like cells (1 nM,83 ％; 10 nM,73 ％; 100 nM,100 ％).Akt levels were higher in BMSCs cultural medium with FCS. Conclusions FCS could induce the differentiation of BMSCs into the cardiomyocyte-like cells. TGF-β1 and BMP-2 might not play a role in the differentiation of BMSCs induced by FCS.BIO enhanced the effects of FCS on the differentiation of BMSCs into cardiomyocyte-like cells,which might involve the Akt pathway.