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1.
Methods Mol Biol ; 1816: 55-66, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29987810

RESUMO

In vitro generated mammalian cardiomyocytes provide experimental models for studying normal mammalian cardiomyocyte development, for disease modeling and for drug development. They also promise to inform future therapeutic strategies for repair of injured or diseased myocardium. Here we provide reliable protocols for differentiation of mouse embryonic stem cells into functional cardiomyocytes, together with Notes about trouble shooting and optimizing such protocols for specific cell lines.


Assuntos
Técnicas de Cultura de Células/métodos , Diferenciação Celular , Células-Tronco Embrionárias Murinas/citologia , Miócitos Cardíacos/citologia , Animais , Linhagem Celular , Meios de Cultura/metabolismo , Camundongos , Células-Tronco Embrionárias Murinas/metabolismo , Miócitos Cardíacos/metabolismo , Soro/metabolismo
2.
Methods Mol Biol ; 1816: 67-78, 2018.
Artigo em Inglês | MEDLINE | ID: mdl-29987811

RESUMO

In vitro generated human cardiomyocytes hold the ultimate promise for heart patients for repair of injured or diseased myocardium, but they also provide experimental models for studying normal cardiomyocyte development, for disease modeling and for drug development. Here we provide reliable protocols for differentiation of human embryonic stem cells into functional cardiomyocytes, together with Notes about troubleshooting and optimizing such protocols for specific cell lines. This chapter also briefly discusses other published protocols and those further adapted for differentiation of induced pluripotent stem cells into cardiomyocytes.


Assuntos
Técnicas de Cultura de Células/métodos , Diferenciação Celular , Células-Tronco Embrionárias Humanas/citologia , Miócitos Cardíacos/citologia , Linhagem Celular , Humanos , Células-Tronco Pluripotentes Induzidas/citologia
3.
Data Brief ; 17: 559-563, 2018 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-29876429

RESUMO

The transcription factors GATA4, GATA5 and GATA6 play important roles in heart muscle differentiation. The data presented in this article are related to the research article entitled "Genome-wide transcriptomics analysis identifies sox7 and sox18 as specifically regulated by gata4 in cardiomyogenesis" (Afouda et al., 2017) [1]. The present study identifies genes regulated by these individual cardiogenic GATA factors using genome-wide transcriptomics analysis. We have presented genes that are specifically regulated by each of them, as well those regulated by either of them. The gene ontology terms (GO) associated with the genes differentially affected are also presented. The data set will allow further investigations on the gene regulatory network downstream of individual cardiogenic GATA factors during cardiac muscle formation.

4.
Dev Biol ; 434(1): 108-120, 2018 02 01.
Artigo em Inglês | MEDLINE | ID: mdl-29229250

RESUMO

The transcription factors GATA4, GATA5 and GATA6 are important regulators of heart muscle differentiation (cardiomyogenesis), which function in a partially redundant manner. We identified genes specifically regulated by individual cardiogenic GATA factors in a genome-wide transcriptomics analysis. The genes regulated by gata4 are particularly interesting because GATA4 is able to induce differentiation of beating cardiomyocytes in Xenopus and in mammalian systems. Among the specifically gata4-regulated transcripts we identified two SoxF family members, sox7 and sox18. Experimental reinstatement of gata4 restores sox7 and sox18 expression, and loss of cardiomyocyte differentiation due to gata4 knockdown is partially restored by reinstating sox7 or sox18 expression, while (as previously reported) knockdown of sox7 or sox18 interferes with heart muscle formation. In order to test for conservation in mammalian cardiomyogenesis, we confirmed in mouse embryonic stem cells (ESCs) undergoing cardiomyogenesis that knockdown of Gata4 leads to reduced Sox7 (and Sox18) expression and that Gata4 is also uniquely capable of promptly inducing Sox7 expression. Taken together, we identify an important and conserved gene regulatory axis from gata4 to the SoxF paralogs sox7 and sox18 and further to heart muscle cell differentiation.


Assuntos
Fator de Transcrição GATA4/metabolismo , Coração/embriologia , Miócitos Cardíacos/metabolismo , Organogênese/fisiologia , Fatores de Transcrição SOXF/biossíntese , Proteínas de Xenopus/biossíntese , Proteínas de Xenopus/metabolismo , Animais , Fator de Transcrição GATA4/genética , Perfilação da Expressão Gênica , Estudo de Associação Genômica Ampla , Camundongos , Células-Tronco Embrionárias Murinas/citologia , Células-Tronco Embrionárias Murinas/metabolismo , Miócitos Cardíacos/citologia , Fatores de Transcrição SOXF/genética , Proteínas de Xenopus/genética , Xenopus laevis
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