Combination of BMP‐2 and 5‐AZA is advantageous in rat bone marrow‐derived mesenchymal stem cells differentiation into cardiomyocytes

Abstract

Bone morphogenetic protein-2 (BMP-2) has a crucial role in the development of cardiogenesis, and is used in inducing bone marrow mesenchymal stem cells (BMMSCs) to differentiate into cardiomyocytes. We have examined a combination of BMP-2 and 5-azacytidine (5-AZA) in inducing these differentiation effects. BMMSCs were collected and purified from bone marrow of 4-week-old Sprague-Dawley (SD) rats by density-gradient centrifugation and differential attachment. The fourth passage subculture of BMMSCs, selected by cytometry for purity and identification, was divided into four groups: a control group, BMP-2 treated, 5-AZA treated, and a combination of BMP-2 and 5-AZA treatment. Expression of cardiac Troponin I (cTnI) and Connexin 43 (CX-43) in BMMSCs after induction were detected by immunofluorescence and Western blot. Flow cytometry analysis was used for differentiation rates and apoptosis of induced BMMSCs, through the expression of cardiac Troponin T (cTnT) and Annexin V-FITC & PI kit, respectively. BMP-2 can ameliorate apoptosis of BMMSCs caused by 5-AZA and promote the differentiation of BMMSCs into cardiomyocyte-like cells. Thus a combination of BMP-2 and 5-AZA can significantly improve the cardiac differentiation with fewer cell damage effects, making it a safe and effective method of induction in vitro.

BMP Signaling Mediated by BMPR1A in Osteoclasts Negatively Regulates Osteoblast Mineralization Through Suppression of Cx43

“…It is reported that BMP‐2 induces expression of Cx43 / Gja1 in cardiomyocyte‐like cells [Degeorge et al, ; Hou et al, ], in an in vitro mesenchymal cell condensation and chondrogenic differentiation model [Zhang et al, ], and in an embryonal mouse P19 cell line [Bani‐Yaghoub et al, ]. While, in immortalized human granulosa cells, theca cell‐derived BMP4 and BMP7 or oocyte‐derived BMP15 down‐regulates Cx43 / Gja1 expression and decreases GJIC activity [Chang et al, , ].…”

Fetal heart extract facilitates the differentiation of human umbilical cord blood-derived mesenchymal stem cells into heart muscle precursor cells

“…In addition, according to some recent publications, the combination of Aza with BMP2 and Wnt1, the combination of Aza and Angiotensin II, or co-culturing stem cells with myocardial cells in inducing medium with Wnt1 or BMP2 has resulted in enhanced differentiation of stem cells into myocardial cells when compared to using each ingredient separately (Hou et al 2013;Xing et al 2012;Zhang et al 2012). Consistent with these reports, our study also showed that the addition of mouse fetal HE into Aza induction medium promoted the expression of cardiomyocyte-specific genes.…”

Promising Therapeutic Strategies for Mesenchymal Stem Cell-Based Cardiovascular Regeneration: From Cell Priming to Tissue Engineering

“…As a conventional inductor, 5-azacytidine (5-AZA) has been used to induce MSC differentiation into cardiomyocytes [53, 93, 94]. Similarly, BMP-2 present in the embryonic heart was used to differentiate ESCs or induce pluripotent stem cells (iPS) into cardiomyocyte-like cells [95, 96], which opens up new possibilities for cardiomyocyte differentiation from MSCs [97]. …”