Differentiation of Bone Marrow Mesenchymal Stem Cells to Cardiomyocyte‐Like Cells Is Regulated by the Combined Low Dose Treatment of Transforming Growth Factor‐β1 and 5‐Azacytidine

Abstract

Bone marrow mesenchymal stem cells (BMMSCs) are used in cardiac tissue engineering for the regeneration of diseased hearts. We examined the differentiation of rat BMMSCs into cardiomyocyte-like cells when induced with a combined low dose treatment of transforming growth factor-β1 (TGF-β1) and 5-azacytidine (5-AZA). Results showed that cell proliferation in the combined low dose treatment group of TGF-β1 and 5-AZA was increased compared with the TGF-β1 group or the 5-AZA group. The cell apoptosis was relieved by combined TGF-β1 and 5-AZA treatment compared to 5-AZA treatment alone. The number of cells positive for myosin heavy chain, connexin-43, α-actin, and troponin I in the combined treatment group was higher than those observed in the TGF-β1 group or the 5-AZA group. Moreover, the combined low dose treatment group of TGF-β1 and 5-AZA reveals the strongest expression of troponin I, α-actin, and phosphorylated extracellular signal-regulated protein kinases 1 and 2 (p-ErK1/2) among the treatment groups. These results suggest that the combined low dose treatment of TGF-β1 and 5-AZA can improve the differentiation potential of rat BMMSCs into cardiomyocyte-like cells and alleviate cell damage effects in vitro. The mechanism that is involved in influencing differentiation may be associated with p-ErK1/2.

Investigation of survival and migration potential of differentiated cardiomyocytes transplanted with decellularized heart scaffold

“…Within this study, we used 5‐Aza for cardiomyogenic gene profile of differentiated AdMSCs into cardiomyocytes. As a result, unlike Shi et al study, our results showed high level of GJA‐1 gene expression which is an important gap junction protein in heart development. Unlike GJA‐1 gene other tested genes did not showed any significant differences between groups .…”

SET domain-containing protein 4: biological functions and role in genomic methylation profiles of bone marrow mesenchymal stem cells

“…We used Transforming growth factor b1 (TGF-b1) plus 5-Aza-2'-deoxycytidine (5-Aza) to induce myogenic differentiation [12]. For myogenic differentiation, 1×10 5 cells were seeded on 6-well plates, and then cells were treated with combination of TGF-b1 (5 ng/mL, #100-21, PeproTech, USA) plus 5-Aza (5 μmol/L, #11390, Sigma-Aldrich, Germany) for 24 h under normoxic condition (21% O 2 ), then medium was changed for DMEM supplemented with 10% FBS (Gibco), TGF-b1 for continuous 14 days.…”

“…The ideal effect of stem cell transplantation for tissue damage depends on the cell transplantation rate, differentiation, and paracrine protective factors. Epigenetics in mesenchymal stem cell differentiation has attracted considerable attention, and most investigations have been focused on osteogenic, adipogenic and cardiogenic differentiation [10][11][12], highlighting epigenetic modification in stem cell biology and regenerative medicine [13].…”

Oxidized low‐density lipoprotein (ox‐LDL) promotes cardiac differentiation of bone marrow mesenchymal stem cells via activating ERK1/2 signaling

“…Blockade of ERK1/2 with the specific inhibitor U0126 could inhibit 5‐Aza‐induced expression of cardiac‐specific genes and proteins in hucMSCs . A recent study showed that the differentiation of BM‐MSCs to cardiomyocyte‐like cells driven by 5‐Aza alone or the combination of 5‐Aza and TGF‐β1 was also through activation of ERK1/2 signaling . Other studies have also shown that the activation of ERK1/2 contributes to cardiac differentiation of human and mouse embryonic stem cells …”

“…7 A recent study showed that the differentiation of BM-MSCs to cardiomyocyte-like cells driven by 5-Aza alone or the combination of 5-Aza and TGF-β1 was also through activation of ERK1/2 signaling. 8 Other studies have also shown that the activation of ERK1/2 contributes to cardiac differentiation of human and mouse embryonic stem cells. [9][10][11] Many different stimuli, including growth factors, cytokines, transforming agents, and oxidative stress can activate ERK signaling.…”