Temporal impact of substrate mechanics on differentiation of human embryonic stem cells to cardiomyocytes.
Acta Biomater
; 10(2): 604-12, 2014 Feb.
Article
em En
| MEDLINE
| ID: mdl-24200714
ABSTRACT
A significant clinical need exists to differentiate human pluripotent stem cells (hPSCs) into cardiomyocytes, enabling tissue modeling for in vitro discovery of new drugs or cell-based therapies for heart repair in vivo. Chemical and mechanical microenvironmental factors are known to impact the efficiency of stem cell differentiation, but cardiac differentiation protocols in hPSCs are typically performed on rigid tissue culture polystyrene (TCPS) surfaces, which do not present a physiological mechanical setting. To investigate the temporal effects of mechanics on cardiac differentiation, we cultured human embryonic stem cells (hESCs) and their derivatives on polyacrylamide hydrogel substrates with a physiologically relevant range of stiffnesses. In directed differentiation and embryoid body culture systems, differentiation of hESCs to cardiac troponin T-expressing (cTnT+) cardiomyocytes peaked on hydrogels of intermediate stiffness. Brachyury expression also peaked on intermediate stiffness hydrogels at day 1 of directed differentiation, suggesting that stiffness impacted the initial differentiation trajectory of hESCs to mesendoderm. To investigate the impact of substrate mechanics during cardiac specification of mesodermal progenitors, we initiated directed cardiomyocyte differentiation on TCPS and transferred cells to hydrogels at the Nkx2.5/Isl1+ cardiac progenitor cell stage. No differences in cardiomyocyte purity with stiffness were observed on day 15. These experiments indicate that differentiation of hESCs is sensitive to substrate mechanics at early stages of mesodermal induction, and proper application of substrate mechanics can increase the propensity of hESCs to differentiate to cardiomyocytes.
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Texto completo:
1
Bases de dados:
MEDLINE
Assunto principal:
Resinas Acrílicas
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Diferenciação Celular
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Miócitos Cardíacos
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Células-Tronco Embrionárias
Tipo de estudo:
Guideline
/
Prognostic_studies
Limite:
Animals
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Humans
Idioma:
En
Revista:
Acta Biomater
Ano de publicação:
2014
Tipo de documento:
Article
País de afiliação:
Estados Unidos