Pluripotent mouse embryonic stem cells are able to differentiate into cardiomyocytes expressing chronotropic responses to adrenergic and cholinergic agents and Ca2+ channel blockers

Wobus, Anna M.; Wallukat, Gerd; Hescheler, Jürgen

doi:10.1111/j.1432-0436.1991.tb00255.x

Cited by 447 publications

(259 citation statements)

References 39 publications

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“…5-Azacytidine interferes with DNA methylation and was shown to induce mouse 10T1/2 fibroblasts to differentiate into skeletal myoblasts by reactivation of the transcription of silenced genes including MyoD [7,8]. After treatment with the drug, immortalized murine bone marrow-derived stromal cells differentiated into cardiomyocytes of various excitation properties [9] similar to the results obtained in cardiomyocytes derived from multipotent embryonic stem cells [10][11][12]. However, conflicting results have been published with respect to MSC from other species.…”

Section: Introductionsupporting

confidence: 55%

5-Azacytidine induces changes in electrophysiological properties of human mesenchymal stem cells

et al. 2006

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Previously, mouse bone marrow-derived stem cells (MSC) treated with the unspecific DNA methyltransferase inhibitor 5-azacytidine were reported to differentiate into cardiomyocytes. The aim of the present study was to investigate the efficiency of a similar differentiation strategy in human mononuclear cells obtained from healthy bone marrow donors. After 1-3 passages, cultures were exposed for 24 h to 5-azacytidine (3 µM) followed by 6 weeks of further culture. Drug treatment did not induce expression of myogenic marker MyoD or cardiac markers Nkx2.5 and GATA-4 and did not yield beating cells during follow-up. In patch clamp experiments, approximately 10-15% of treated and untreated cells exhibited L-type Ca 2+ currents. Almost all cells showed outwardly rectifying K + currents of rapid or slow activation kinetics. Mean current amplitude at +60 mV doubled after 6 weeks of treatment compared with time-matched controls. Membrane capacitance of treated cells was significantly larger than in controls 2 weeks after treatment and remained high after 6 weeks. Expression levels of mRNAs for the K + channels Kv1.1, Kv1.5, Kv2.1, Kv4.3 and KCNMA1 and for the Ca 2+ channel Ca v 1.2 were not affected by 5-azacytidine. Treatment with potassium channel blockers tetraethylammonium and clofilium at concentrations shown previously to inhibit rapid or slowly activating K + currents of hMSC inhibited proliferation of these cells. Our results suggest that despite the absence of differentiation of hMSC into cardiomyocytes, treatment with 5-azacytidine caused profound changes in current density.Cell Research (2006) 16:949-960.

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Section: Introductionsupporting

confidence: 55%

5-Azacytidine induces changes in electrophysiological properties of human mesenchymal stem cells

et al. 2006

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“…Despite extended periods in culture, ES cells retain the capacity to differentiate into all fetal and adult cell types [3], a characteristic best exemplified by the ability of murine ES cells to integrate into host blastocysts and contribute to germ line and somatic tissue in chimeras [4]. ES cells can also originate a variety of specialized cell lineages in vitro, after aggregation into three-dimensional structures termed embryoid bodies (EBs) [5], and can provide a powerful model system to study mechanisms and genetic circuits involved in the earliest steps of embryonic development and lineage specification [6]. It is evident that the exploitation of EB differentiation to defined progenitor types could potentially provide an unlimited source of functional replacement cells for therapeutic interventions [7].…”

Section: Introductionmentioning

confidence: 99%

Mitochondrial Reactive Oxygen Species Mediate Cardiomyocyte Formation from Embryonic Stem Cells in High Glucose

et al. 2010

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Accumulating evidence points to reactive oxygen species (ROS) as important signaling molecules for cardiomyocyte differentiation in embryonic stem (ES) cells. Given that ES cells are normally maintained and differentiated in medium containing supraphysiological levels of glucose (25 mM), a condition which is known to result in enhanced cellular ROS formation, we questioned whether this high glucose concentration was necessary for cardiomyocyte lineage potential. We show here that ES cells cultured in physiological glucose (5 mM), maintained their general stemness qualities but displayed an altered mitochondrial metabolism, which resulted in decreased ROS production. Furthermore, ES and induced pluripotent stem (iPS) cells differentiated in lower glucose concentrations failed to generate cardiomyocyte structures; an effect mimicked with antioxidant treatments using catalase, N-acetyl cysteine and mitoubiquinone, under high glucose conditions in ES cells. Molecular analysis revealed that ES cells differentiated in 5 mM glucose had reduced expression of the pro-cardiac NOX4 gene and diminished phosphorylation of p38 mitogen-activated protein kinase (MAPK), together with specific changes in the cardiac transcriptional network. These outcomes could be reversed by supplementation of low glucose cultures with ascorbic acid, paradoxically acting as a pro-oxidant. Furthermore, forced expression of an upstream p38 MAPK kinase (MKK6) could bypass the requirement for ROS during differentiation to cardiomyocytes under low glucose conditions, illustrating a key role for p38 in the cardiac differentiation program. Together these data demonstrate that endogenous ROS control is important for cardiomyocyte formation from ES cells, and furthermore that supraphysiological glucose, by supplying ROS, is absolutely required.

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“…Embryoid body (EB) induction in the suspension culture (Stanford et al, 1998) or the hanging drop culture (Wobus et al, 1991) was carried out essentially as described, except that 30-l drops containing 800 -1,000 cells were set up and that standard ES cell medium was used. Resulting EBs were transferred in groups (if formed in the suspension culture) or individually (if formed in the hanging drop culture) into gelatin-coated 24-well plates on day 4 for subsequent attachment cultures.…”

Section: In Vitro Differentiationmentioning

confidence: 99%

“…In addition to the specialized structures they form, these cells exhibit characteristic activities that are hallmarks for lineage identification. The most striking activities are the spontaneous, rhythmical contractions of cardiomyocyte aggregates (Wobus et al, 1991). Smooth muscle cells are found in tightly packed bundles of thin fibers showing slow, wave-like contractions (Weitzer et al, 1995;Drab et al, 1997), which differ markedly from the much thicker, fastcontracting skeletal muscle myotubes (Rohwedel et al, 1994).…”

Section: Introductionmentioning

confidence: 99%

Differentiation trapping screen in live culture for genes expressed in cardiovascular lineages

Chen

2004

Developmental Dynamics

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Pluripotent mouse embryonic stem cells are able to differentiate into cardiomyocytes expressing chronotropic responses to adrenergic and cholinergic agents and Ca2+ channel blockers

Cited by 447 publications

References 39 publications

5-Azacytidine induces changes in electrophysiological properties of human mesenchymal stem cells

5-Azacytidine induces changes in electrophysiological properties of human mesenchymal stem cells

Mitochondrial Reactive Oxygen Species Mediate Cardiomyocyte Formation from Embryonic Stem Cells in High Glucose

Differentiation trapping screen in live culture for genes expressed in cardiovascular lineages

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