Temporal impact of substrate mechanics on differentiation of human embryonic stem cells to cardiomyocytes

Laurie B. Hazeltine, Mehmet G. Badur, Xiaojun Lian, Amritava Das, Wenqing Han, Sean P. Palecek

Research output: Contribution to journalArticle

39 Citations (Scopus)

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.

Original languageEnglish (US)
Pages (from-to)604-612
Number of pages9
JournalActa Biomaterialia
Volume10
Issue number2
DOIs
StatePublished - Feb 2014

Fingerprint

Stem cells
Mechanics
Cardiac Myocytes
Hydrogels
Substrates
Pluripotent Stem Cells
Stiffness
Polystyrenes
Stem Cells
Tissue culture
Embryoid Bodies
Troponin T
Drug Discovery
Cell- and Tissue-Based Therapy
Cell Differentiation
Human Embryonic Stem Cells
Polyacrylates
Cell culture
Repair
Trajectories

All Science Journal Classification (ASJC) codes

  • Biotechnology
  • Biomaterials
  • Biochemistry
  • Biomedical Engineering
  • Molecular Biology

Cite this

Hazeltine, Laurie B. ; Badur, Mehmet G. ; Lian, Xiaojun ; Das, Amritava ; Han, Wenqing ; Palecek, Sean P. / Temporal impact of substrate mechanics on differentiation of human embryonic stem cells to cardiomyocytes. In: Acta Biomaterialia. 2014 ; Vol. 10, No. 2. pp. 604-612.
@article{1150bbf40651448d82f5edcc4df364f1,
title = "Temporal impact of substrate mechanics on differentiation of human embryonic stem cells to cardiomyocytes",
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.",
author = "Hazeltine, {Laurie B.} and Badur, {Mehmet G.} and Xiaojun Lian and Amritava Das and Wenqing Han and Palecek, {Sean P.}",
year = "2014",
month = "2",
doi = "10.1016/j.actbio.2013.10.033",
language = "English (US)",
volume = "10",
pages = "604--612",
journal = "Acta Biomaterialia",
issn = "1742-7061",
publisher = "Elsevier BV",
number = "2",

}

Temporal impact of substrate mechanics on differentiation of human embryonic stem cells to cardiomyocytes. / Hazeltine, Laurie B.; Badur, Mehmet G.; Lian, Xiaojun; Das, Amritava; Han, Wenqing; Palecek, Sean P.

In: Acta Biomaterialia, Vol. 10, No. 2, 02.2014, p. 604-612.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Temporal impact of substrate mechanics on differentiation of human embryonic stem cells to cardiomyocytes

AU - Hazeltine, Laurie B.

AU - Badur, Mehmet G.

AU - Lian, Xiaojun

AU - Das, Amritava

AU - Han, Wenqing

AU - Palecek, Sean P.

PY - 2014/2

Y1 - 2014/2

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84896544312&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84896544312&partnerID=8YFLogxK

U2 - 10.1016/j.actbio.2013.10.033

DO - 10.1016/j.actbio.2013.10.033

M3 - Article

C2 - 24200714

AN - SCOPUS:84896544312

VL - 10

SP - 604

EP - 612

JO - Acta Biomaterialia

JF - Acta Biomaterialia

SN - 1742-7061

IS - 2

ER -