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Stem Cell Mechanosensation on Gelatin Methacryloyl (GelMA) Stiffness Gradient Hydrogels

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Abstract

Stiffness gradient hydrogels are a useful platform for studying mechanical interactions between cells and their surrounding environments. Here, we developed linear stiffness gradient hydrogels by controlling the polymerization of gelatin methacryloyl (GelMA) via differential UV penetration with a gradient photomask. Based on previous observations, a stiffness gradient GelMA hydrogel was created ranging from ~ 4 to 13 kPa over 15 mm (0.68 kPa/mm), covering the range of physiological tissue stiffness from fat to muscle, thereby allowing us to study stem cell mechanosensation and differentiation. Adipose-derived stem cells on these gradient hydrogels showed no durotaxis, which allowed for the screening of mechanomarker expression without confounding directed migration effects. In terms of morphological markers, the cell aspect ratio showed a clear positive correlation to the underlying substrate stiffness, while no significant correlation was found in cell size, nuclear size, or nuclear aspect ratio. Conversely, expression of mechanomarkers (i.e., Lamin A, YAP, and MRTFa) all showed a highly significant correlation to stiffness, which could be disrupted via inhibition of non-muscle myosin or Rho/ROCK signalling. Furthermore, we showed that cells plated on stiffer regions became stiffer themselves, and that stem cells showed stiffness-dependent differentiation to fat or muscle as has been previously reported in the literature.

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Acknowledgments

This study work was supported by National Health and Medical Research Council Grant PG1098449 (to YSC), Heart Foundation Future Leader Fellowship 101173 (to YSC), Department of Health, Western Australia, Merit awards—Project and fellowship (to YSC), and Universities Australia DAAD German Research Cooperation 5744610 (to YSC, CK, AWH, JLY and JPS).

Author information

Authors and Affiliations

  1. School of Human Sciences, The University of Western Australia, 35 Stirling Highway, Crawley, WA, 6009, Australia

    Claire Kim, Luke G. Major & Yu Suk Choi

  2. Department of Cellular Biophysics, Max Planck Institute for Medical Research, 69120, Heidelberg, Germany

    Jennifer L. Young, Andrew W. Holle & Joachim P. Spatz

  3. Department of Biophysical Chemistry, University of Heidelberg, 69117, Heidelberg, Germany

    Jennifer L. Young, Andrew W. Holle & Joachim P. Spatz

  4. Fluid Science and Resources, Department of Chemical Engineering, School of Engineering, University of Western Australia, Perth, WA, 6009, Australia

    Kwanghee Jeong & Zachary M. Aman

  5. Soonchunhyang Institute of Medi-bio Science, Soonchunhyang University, Cheonan-si, Chungcheongnam-do, 31151, Korea

    Ji Hoon Jeong & Yongsung Hwang

  6. Department of CognoMechatronics Engineering, College of Nanoscience & Nanotechnology, Pusan National University, Busan, 46241, Korea

    Dong-Wook Han

Authors
  1. Claire Kim
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Contributions

CK, LGM and YSC designed and planned the study. JLY, AWH, and JPS performed SEM and pore size analysis. KJ and ZMA performed Raman spectroscopy. JHJ, YH and DWH synthesized GelMA. CK, JLY and YSC drafted the manuscript. YSC provided supervision and funding. All authors discussed the data and contributed to the final version of the manuscript.

Corresponding author

Correspondence to Yu Suk Choi.

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Associate Editor Kent Leach oversaw the review of this article.

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Kim, C., Young, J.L., Holle, A.W. et al. Stem Cell Mechanosensation on Gelatin Methacryloyl (GelMA) Stiffness Gradient Hydrogels. Ann Biomed Eng 48, 893–902 (2020). https://doi.org/10.1007/s10439-019-02428-5

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  • DOI: https://doi.org/10.1007/s10439-019-02428-5

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