[Wnt3a combined with bone morphogenetic protein 9 induce C3H10T1/2 cells differentiation into cardiomyocyte-like cells in vitro].

OBJECTIVE: To investigate the effects of Wnt3a alone and combined with bone morphogenetic protein 9 (BMP9) on the differentiation of C3H10T1/2 cells into cardiomyocyte-like cells.

METHODS: Recombinant adenovirus GFP, BMP9 and Wnt3a were amplified with human embryo kidney 293 cell line (HEK293 cells) and transferred into C3H10T1/2 cells. Three weeks after transfection, the cell morphology and the expression of green fluorescent protein in cells transfected with GFP, BMP9 and Wnt3a were observed by an inverted microscope and a fluorescence microscope. Flow cytometry was performed to detect cell transfection efficiency. The expressions of cardiac-specific proteins connexin 43 (Cx43), cardiac troponin T (cTnT) and genes GATA binding protein 4 (GATA4), myocyte enhancer factor 2C (MEF2C) were analyzed by Western blotting, immunofluorescence and real-time quantitative PCR (qRT-PCR).

RESULTS: High-titer recombinant adenovirus was generated with HEK293 cells. The expressions of Cx43, cTnT, GATA4 and MEF2C were similar among Wnt3a group, GFP group and control group, but they significantly increased in cells when co-induced by Wnt3a and BMP9 for 3 weeks. The expressions of Cx43, cTnT, GATA4 and MEF2C showed no significant difference between BMP9 group and Wnt3a and BMP9 co-induction group.

CONCLUSION: Wnt3a can not promote the differentiation of C3H10T1/2 cells into cardiomyocyte-like cells by itself, but co-induction by Wnt3a and BMP9 can promote the differentiation.