Stiffness‐Tunable Hydrogel‐Sandwich Culture Modulates the YAP‐Mediated Mechanoresponse in Induced‐Pluripotent Stem Cell Embryoid Bodies and Augments Cardiomyocyte Differentiation

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  • Praphawi Nattasit
    Division of Molecular and Regenerative Prosthodontics Tohoku University Graduate School of Dentistry Sendai Miyagi 980‐8575 Japan
  • Kunimichi Niibe
    Division of Molecular and Regenerative Prosthodontics Tohoku University Graduate School of Dentistry Sendai Miyagi 980‐8575 Japan
  • Masahiro Yamada
    Division of Molecular and Regenerative Prosthodontics Tohoku University Graduate School of Dentistry Sendai Miyagi 980‐8575 Japan
  • Yumi Ohori‐Morita
    Division of Molecular and Regenerative Prosthodontics Tohoku University Graduate School of Dentistry Sendai Miyagi 980‐8575 Japan
  • Phoonsuk Limraksasin
    Division of Molecular and Regenerative Prosthodontics Tohoku University Graduate School of Dentistry Sendai Miyagi 980‐8575 Japan
  • Watcharaphol Tiskratok
    Division of Molecular and Regenerative Prosthodontics Tohoku University Graduate School of Dentistry Sendai Miyagi 980‐8575 Japan
  • Masaya Yamamoto
    Department of Material Processing Tohoku University Graduate School of Engineering Sendai Miyagi 980‐8579 Japan
  • Hiroshi Egusa
    Division of Molecular and Regenerative Prosthodontics Tohoku University Graduate School of Dentistry Sendai Miyagi 980‐8575 Japan

説明

<jats:title>Abstract</jats:title><jats:p>Microenvironmental factors, including substrate stiffness, regulate stem cell behavior and differentiation. However, the effects of substrate stiffness on the behavior of induced pluripotent stem cell (iPSC)‐ derived embryoid bodies (EB) remain unclear. To investigate the effects of mechanical cues on iPSC‐EB differentiation, a 3D hydrogel‐sandwich culture (HGSC) system is developed that controls the microenvironment surrounding iPSC‐EBs using a stiffness‐tunable polyacrylamide hydrogel assembly. Mouse iPSC‐EBs are seeded between upper and lower polyacrylamide hydrogels of differing stiffness (Young's modulus [E’] = 54.3 ± 7.1 kPa [hard], 28.1 ± 2.3 kPa [moderate], and 5.1 ± 0.1 kPa [soft]) and cultured for 2 days. HGSC induces stiffness‐dependent activation of the yes‐associated protein (YAP) mechanotransducer and actin cytoskeleton rearrangement in the iPSC‐EBs. Moreover, moderate‐stiffness HGSC specifically upregulates the mRNA and protein expression of ectoderm and mesoderm lineage differentiation markers in iPSC‐EBs via YAP‐mediated mechanotransduction. Pretreatment of mouse iPSC‐EBs with moderate‐stiffness HGSC promotes cardiomyocyte (CM) differentiation and structural maturation of myofibrils. The proposed HGSC system provides a viable platform for investigating the role of mechanical cues on the pluripotency and differentiation of iPSCs that can be beneficial for research into tissue regeneration and engineering.</jats:p>

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