5-Azacytidine-Induced Cardiomyocyte Differentiation of Very Small Embryonic-Like Stem Cells

Abstract

The use of stem cells in generating cell-based pacemaker therapies for bradyarrhythmia is currently being considered. Due to the propensity of stem cells to form tumors, as well as ethical issues surrounding their use, the seed cells used in cardiac biological pacemakers have limitations. Very small embryonic-like stem cells (VSELs) are a unique and rare adult stem cell population, which have the same structural, genetic, biochemical, and functional characteristics as embryonic stem cells without the ethical controversy. In this study, we investigated the ability of rat bone marrow- (BM-) derived VSELs to differentiate in vitro into cardiomyocytes by 5-Azacytidine (5-AzaC) treatment. The morphology of VSELs treated with 10 μM 5-AzaC increased in volume and gradually changed to cardiomyocyte-like morphology without massive cell death. Additionally, mRNA expression of the cardiomyocyte markers cardiac troponin-T (cTnT) and α-sarcomeric actin (α-actin) was significantly upregulated after 5-AzaC treatment. Conversely, stem cell markers such as Nanog, Oct-4, and Sox2 were continuously downregulated posttreatment. On day 14 post-5-AzaC treatment, the positive expression rates of cTnT and α-actin were 18.41±1.51% and 19.43±0.51%, respectively. Taken together, our results showed that rat BM-VSELs have the ability to differentiate into cardiomyocytes in vitro. These findings suggest that VSELs would be useful as seed cells in exploring the mechanism of biological pacemaker activity.

The roles of DNA methylation on the promotor of the Epstein–Barr virus (EBV) gene and the genome in patients with EBV-associated diseases

“…5-AzaC, a demethylating drug, mainly binds to DNA methyltransferase through covalent bonds to form an irreversible complex, which reduces the level of genomic methylation and reactivates hypermethylated tumour suppressor genes. At present, many studies have confirmed that most of the CpG islands in the promoter region of many tumour suppressor genes are in a state of hypermethylation, and the expression of protein can be restored after intervention with the demethylated drug 5-AzaC (Sun et al 2020 ). Additionally, it has been reported that zebularine can activate the tumor suppressor genes p15 , p16 , and p57 by demethylation in gastric cancer cells (Pan et al 2016 ).…”

H3K9me2 regulates early transcription factors to promote mesenchymal stem‑cell differentiation into cardiomyocytes

“…Currently, the efficiency of stem cell induction into cardiomyocytes is low, which limits its clinical application. Our previous study treated very small embryonic-like stem cells with 5-AZA and the positive expression rates of cTnT and α-actin were 18.41 and 19.43%, respectively on day 14 (19). In the present study, epigenetics was used to improve the differentiation efficiency of stem cells.…”

Histone demethylase KDM5B catalyzed H3K4me3 demethylation to promote differentiation of bone marrow mesenchymal stem cells into cardiomyocytes

“…There are some differences between these two kinds of cells in terms of extraction methods, culture conditions and morphological characteristics. The morphological changes during induction differentiation were not identical [27,28].…”

“…The mechanism of 5-AZA may be related to CpG base-pair demethylation and regulation of early myocardial transcription factors [32,33]. Currently, the efficiency of stem cell induction into cardiomyocytes is low, which limits its clinical application [34]. In the present study, epigenetics was used to improve the differentiation efficiency of stem cells.…”