Enhancement of cardiomyocyte differentiation from human embryonic stem cells

Abstract

Several approaches have been used to encourage the differentiation of cardiomyocytes from human embryonic stem cells. However, the differentiation efficiency is low, and appropriate culture protocols are needed to produce adequate numbers of cardiomyocytes for therapeutic cell transplantation. This study investigated the effects of serum on cardiomyocyte differentiation in suspension culture medium during embryoid body (EB) formation by human embryonic stem cells. The addition of ascorbic acid, dimethylsulfoxide and 5-aza-2'-deoxycytidine during days 5-7 at the EB-forming stage resulted in an increase in the numbers of rhythmically contracting clusters of derived cardiomyocytes. Treatment with 0.1 mmol L(-1) ascorbic acid alone, or more notably in combination with 10 micromol L(-1) 5-aza-2'-deoxycytidine, induced the formation of beating cells within EBs. Most of the beating clusters had spontaneous contraction rates similar to those found in human adults, and their contractile activity lasted for up to 194 days.

Diverse effects of dimethyl sulfoxide (DMSO) on the differentiation potential of human embryonic stem cells

“…Evaluation of mesodermal markers like HAND1, MEF2C, GATA-4 and C-actin revealed a considerable upregulation of all these genes in low and medium dose, whereas highest dose of DMSO triggered significant downregulation of HAND1, GATA-4 and C-actin. These results indicate that DMSO at low and medium doses can act as an inducing agent for the formation of mesodermal phenotypes as also evidenced by the earlier reports (Wang et al 2010). Although DMSO is known to differentiate P19CL6 embryonic carcinoma cells into cardiomyocyte-like cells, the low differentiation capacity of DMSO reduces its usefulness (Seya et al 2011).…”

“…3c). Our results reassure the long-standing concept that DMSO at low and medium concentrations can act as an inducing agent for the formation of mesodermal phenotypes (Wang et al 2010).…”

Effects of C-Reactive Protein on the Cardiac Differentiation of Mouse Embryonic Stem Cells

“…There are studies and reports demonstrating that many signaling pathways or compounds are involved in, such as STAT3, BMP, Wnt, FGF, PGC-1α, and reactive oxygen species. [25][26][27][28][29][30] It was not surprising that the state of the signaling pathways or compounds during differentiation might be modulated by a strong extracellular stimulus of 150 mg/L CRP, but not by a weaker stimulus of 30 mg/L CRP. Nevertheless, the present study demonstrated the effect of 150 mg/L CRP on the cardiac differentiation of ES cells.…”

Potential for pharmacological manipulation of human embryonic stem cells

“…NaB has also been shown to promote the rapid differentiation of hESCs to primitive endoderm and trophectoderm lineages induced by nutlin, a small molecule activator of p53 (Maimets et al, 2008). Cardiomyocyte differentiation has been demonstrated to be enhanced by 5-azacytidine/5-aza-2′-deoxycytidine (Xu et al, 2002;Yoon et al, 2006;Wang et al, 2010), a chemical analogue of cytidine that acts as a false substrate for DNA methyltransferases, therefore reducing cellular DNA methylation content. A reduction in DNA methylation, similar to an increase in histone acetylation, induces the reactivation of genes associated with the differentiation of hESCs, and thereby primes them for appropriate signals to allow lineagespecific differentiation.…”