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...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.展开更多
文摘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.
