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2022, Vol. 26 ›› Issue (19): 2958-29635-Azacytidine combined with bone morphogenetic protein 2 induces differentiation of bone marrow mesenchymal stem cells into cardiomyocyte-like cellsWang Wenhua1, 2, Liu Yang2, Wang Qiaomin1, 2, Lü Yang2, Zhao Yaru1, 2, Wang Haiping2
Abstract: BACKGROUND: Stem cells have become ideal seed cells in tissue engineering, which provides a research direction for cell transplantation in the treatment of myocardial infarction. OBJECTIVE: To investigate the feasibility of bone morphogenetic protein 2 combined with 5-azacytidine on the differentiation of bone marrow mesenchymal stem cells into cardiomyocyte-like cells in vitro. METHODS: Bone marrow mesenchymal stem cells were cultured by whole bone marrow culture adherent method. The third generation of bone marrow mesenchymal stem cells was divided into four groups: control group (ordinary Iscove’s modified Dulbecco’s medium), bone morphogenetic protein 2 group (200 ng/L), 5-azacytidine group (10 µmol/L), 5-azacytidine and bone morphogenetic protein 2 group. After induction for 3 days, the normal medium was changed and cultured for 4 weeks. The expression of cTnI was detected by immunocytochemistry. The expression of Desmin and cTnT was detected by immunofluorescence. The expression of cTnT and cTnI was detected by western blot assay. After 3 days of induction, the normal medium was changed and cultured for 1, 2 and 4 weeks. The mRNA expression of GATA-4 and Nkx2.5 was detected by RT-qPCR. RESULTS AND CONCLUSION: (1) Western blot assay showed that cTnT and cTnI were expressed in the induction group. The expression levels of cTnT and cTnI in the combined group were higher than those in other groups (P < 0.01). (2) Immunocytochemistry showed positive expression of cardiac troponin (cTnI) in the induction group; the expression was higher in the combined group than that in the induction group (P < 0.05). Immunofluorescence cytochemistry assay showed the positive expression of Desmin and cTnT; the expression in the combined group was higher than that in the induction group (P < 0.05). (3) RT-qPCR showed the mRNA expression of GATA-4 and Nkx2.5 in the induction group; the expression was higher in the combined group than that in the induction group (P < 0.05). (4) The results showed that bone marrow mesenchymal stem cells induced by 5-azacytidine and bone morphogenetic protein 2 could improve the differentiation efficiency of cardiomyocyte-like cells. Key words:stem cells, bone marrow mesenchymal stem cells, myocardial like cells, bone morphogenetic protein 2, 5-azacytidine, SD rats |