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.

Shutian Shi,Huangtai Miao,Shaoping Nie,Xiao Wang,Wen Hao,Xingxin Wu,Lei Zhen

doi:10.1155/2016/3816256

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.

Highlights

Ischemic heart disease is a leading cause of morbidity and mortality in developed countries and is one of the major contributors of disease burden in developing countries [1, 2]
We investigated the effect of combined low dose treatment of transforming growth factor-β1 (TGF-β1) and 5-AZA, on cardiomyogenic differentiation of Bone marrow-derived mesenchymal stem cells (BMMSCs)
The cells ended up either as polygonal or as long, spindle-shaped cells. These cells were confirmed as mesenchymal stem cells (MSCs) as they stained positive for CD29 (90.26 ± 1.41%), CD44 (93.44 ± 3.65%), and CD90 (98.82 ± 4.03%), while they stained negative for CD19 (3.61 ± 0.37%), CD34 (2.92 ± 0.29%), and CD45 (1.87 ± 0.44%)

Summary

Introduction

Ischemic heart disease is a leading cause of morbidity and mortality in developed countries and is one of the major contributors of disease burden in developing countries [1, 2]. The current treatment techniques that include medications, interventional procedures, and surgery can only open occluded coronary arteries or provide symptomatic relief without addressing the major issue of the cumulative loss of functioning cardiomyocytes [3,4,5]. Cellular therapy offers another option in the treatment of ischemic heart diseases. 5-AZA can affect cell survival after induction because of its obvious toxicity as an antineoplastic agent [10], while TGF-β1 reveals very low efficiency [11]. One of the main challenges of cell therapy is to identify an induction method with a high differentiation rate and low cytotoxicity

Methods

Results

Conclusion