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| Study of sodium butyrate combined with fibroblast growth factor 2 on differentiation of bone marrow mesenchymal stem cells into cardiomyocytes |
| ZHAO Yaru LIU Yang LYU Yang WANG Wenhua WANG Haoyu WANG Qiaomin WANG Haiping |
| Department of Histology and Embryology, Hebei North University, Hebei Province, Zhangjiakou 075000, China |
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Abstract Objective To investigate the feasibility of inducing the differentiation of bone marrow mesenchymal stem cells (BMMSC) into cardiomyocytes by sodium butyrate and fibroblast growth factor 2 (FGF-2) in vitro. Methods BMMSC were isolated and purified by whole bone marrow adherent method and identified by flow cytometry. Normal complete medium (control group), 1 mmol/L sodium butyrate medium (sodium butyrate group), 1 ng/ml FGF-2 medium (FGF-2 group), 1 mmol/L sodium butyrate and 1 ng/ml FGF-2 medium (combined group) were added to induce culture. Total RNA was extracted from four groups at 1, 2 and 4 weeks after induction. Myocardial specific transcription factors were detected by quantitative reverse transcriptase-mediated PCR (qRT-PCR). Immunocytochemistry and Western blot were used to detect the expression of myocardial specific proteins induced by BMMSC after four weeks. Results After 48 h of culture, the primary cells showed oblate and adherent growth, and some cells had short fusiform changes. BMMSC cultured for one week were further extended and showed spindle shape, polygon shape and irregular shape. BMMSC cultured for four weeks showed obvious directional arrangement between adjacent cells, and some cells even formed whirlpool-like structures. Flow cytometry results showed that the isolated cells were purified BMMSC. The results of qRT-PCR showed that the expression of myocardium specific gene GATA-4 was the highest after two weeks of culture, and the expression in the combined group was significantly higher than that in the other groups (P ＜ 0.05). The results of Western blot and immunocytochemistry showed that after four weeks of BMMSC culture, the heart-specific protein level in combined group was higher than those in the other groups (P ＜ 0.05). Conclusion Sodium butyrate and FGF-2 induced the differentiation of BMMSC into cardiomyoid cells, and the combined induction is better than the single induction.
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