Ultrastructural maturation of human bone marrow mesenchymal stem cells‐derived cardiomyocytes under alternative induction of 5‐azacytidine

Abstract

Adult cardiomyocytes lack the ability to proliferate and are unable to repair damaged heart tissue, therefore differentiation of stem cells to cardiomyocytes represents an exceptional opportunity to study cardiomyocytes in vitro and potentially provides a valuable source for replacing damaged tissue. However, characteristic maturity of the in vitro differentiated cardiomyocytes and methods to achieve it are yet to be optimized. In this study, differentiation of human bone marrow-mesenchymal stem cells (hBM-MSCs) into cardiomyocytes is accomplished and the process investigated ultrastructurally. The hBM-MSCs were alternatively treated with 5 μM of 5-azacytidine (5-aza) for 8 weeks resulting in differentiation to cardiomyocytes. Expressions of cardiomyocyte-specific genes [cardiac α-actinin, cardiac β-myosin heavy chain (MHC) and connexin-43] and proteins (cardiac α-actinin, cardiac troponin and connexin-43) were confirmed in a time-dependent manner from the first to the fifth weeks post-induction. Ultrastructural maturation of hBM-MSCs-derived cardiomyocyte (MSCs-CM) corresponded with increase in number and organization of myofilaments in cells over time. Starting from week five, organized myofibrils along with developing sarcomeres were detectable. Later on, MSCs-CM were characterized by the presence of sarcoplasmic reticulum, T-tubules and diads as cardiomyocytes connected to each other by intercalated disc-like structures. Here, we showed the potential of hBM-MSCs as a source for the production of cardiomyocytes and confirmed mature ultrastructural characteristics of these cells using our alternative incubation method.

Cardiac patch design: compatibility of nanofiber materials prepared byelectrospinning method with stem cells

“…These results fit well with previous in vitro cytocompatibility tests carried out on PU fibers (Kuo et al, 2014). It was previously reported that MSCs can differentiate into cardiomyocyte-like cells after 5-azacytidine treatment (Antonitsis et al, 2007;Zhang et al, 2009;Li et al, 2013; Supokawej et al, 2013;Wu et al, 2013;Piryaei et al, 2015). Nartprayut et al showed the ability of perinatally derived MSC differentiation into cardiomyocyte-like cells (after 5-azacytidine treatment) that expressed the cardiac-specific genes Nkx2.5, α-cardiac actin, and cardiac troponin T .…”

Three‐dimensional collagenous niche and azacytidine selectively promote time‐dependent cardiomyogenesis from human bone marrow‐derived MSC spheroids

“…Beating cardiomyogenic cells were formed from murine bone marrow stromal cells after 2 weeks of culture, following treatment with 3 μM aza for 24 hr (Makino et al, 1999). On the other hand, continuous exposure to aza resulted in greater cell death, while single exposure to aza caused relatively smaller fraction of MSCs to differentiate into immature phenotype (Piryaei et al, 2015). On the other hand, continuous exposure to aza resulted in greater cell death, while single exposure to aza caused relatively smaller fraction of MSCs to differentiate into immature phenotype (Piryaei et al, 2015).…”

Cells, Scaffolds and Their Interactions in Myocardial Tissue Regeneration

“…Several pre‐clinical and clinical studies have evaluated some cell types for cardiac regeneration, including skeletal myoblasts, mesenchymal stem cells (bone marrow‐derived and adipose‐derived), embryonic stem cells, and cardiac stem cells. The majority of these cell types are produced preclinically and are safe and effective in clinical practice; however, it has been shown that cardiac stem cells or cardiopoietic stem cells are more effective than other cell types and surgeons are thus keen to use lineage‐specified cells derived from the heart for ideal treatment of heart diseases [Ahmadi et al, ; Piryaei et al, ]. A summary of the findings of studies used in this review is shown in Table .…”