Angiotensinand 5 - azacytidine induced by bone marrow mesenchymal stem cells into cardiomyocyte-like cells.docVIP

 PAGE \* MERGEFORMAT 10 Angiotensinand 5 - azacytidine induced by bone marrow mesenchymal stem cells into cardiomyocyte-like cells Of: Xing Yujie, Lvan Lin, Wang Li 1, Yan Xuebo in the military, IgY [Abstract] Objective To observe the angiotensin in combination with 5 - azacytidine in vitro differentiation of bone marrow mesenchymal stem cells (BMMSCs) differentiate into cardiomyocyte-like cells. Methods of rats were isolated by density gradient centrifugation BMMSCs, passage 3 BMMSCs grouped induced: Angiotensin group (Ang , a final concentration of 0.1mol / L), 5 - azacytidine group (5-aza, a final concentration of 10mol / L), Ang combined 5-aza group (final concentration were 0.1mol / L and 10mol / L), blank control group. induced 24h after the replacement of conventional culture medium and cultured for 4 weeks. inverted phase contrast microscope observation of cell morphological changes, MTT assay and the proliferation of cell activity, immunofluorescence staining Identification of specific induction of myocardial troponin BMMSCs I (cTnI) expression, flow cytometry method to calculate the induction rate of myocardial cells, TEM cells after induction of myocardial ultrastructure. BMMSCs14 primary culture results days to form colonies, passaged cells became larger, elongated spindle cells after induction, showing consistent growth, and the emergence of muscle island-like structure. MTT assay showed that combined 5-aza Ang cell proliferation better than Ang group and 5 - N cytidine group. immunofluorescence staining showed that induced expression of cardiac-specific proteins BMMSCs cTnI. flow cytometry showed: Ang group ,5-aza, Ang combined 5-aza induction rate of myocardial cells were (25.3 + - 2.2)%, (24.6 + -1.9)%, (30.0 + -1.7)%, Ang combined 5-aza group was higher than that of myocardial cells induced by Ang group and 5 - azacytidine group (Pgt; 0.05). Transmission electron microscopy parallel arrangement of filaments can be seen, Z line-li