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ORIGINAL RESEARCH article

Front. Bioeng. Biotechnol.
Sec. Cell and Gene Therapy
Volume 12 - 2024 | doi: 10.3389/fbioe.2024.1467412
This article is part of the Research Topic Advances in Stem Cell Engineering: Paving the Way for Regenerative Medicine View all articles

Morphogen-driven differentiation is precluded by physical confinement in human iPSCs spheroids

Provisionally accepted
Haneen S. Alsehli Haneen S. Alsehli 1,2*Errin Roy Errin Roy 2Thomas Williams Thomas Williams 2Alicja Kuziola Alicja Kuziola 3Yunzhe Guo Yunzhe Guo 3Jeremy B. Green Jeremy B. Green 3Eileen Gentleman Eileen Gentleman 3,4Davide Danovi Davide Danovi 2,5*
  • 1 Centre for Stem Cell Biology, The University of Sheffield, Sheffield, United Kingdom
  • 2 Centre for Stem Cells & Regenerative Medicine, School of Basic & Medical Biosciences, Faculty of Life Sciences & Medicine, King's College London, London, England, United Kingdom
  • 3 Centre for Craniofacial and Regenerative Biology, Faculty of Dentistry, Oral & Craniofacial Sciences, King’s College London, London, England, United Kingdom
  • 4 Department of Biomedical Sciences, Faculty of Biology and Medicine, University of Lausanne, Lausanne, Vaud, Switzerland
  • 5 bit.bio, Babraham Research Campus, Cambridge, United Kingdom

The final, formatted version of the article will be published soon.

    Cell lineage specification is tightly associated with profound morphological changes in the developing human embryo, particularly during gastrulation. The interplay between mechanical forces and biochemical signals is poorly understood. Here, we dissect the effects of biochemical cues and physical confinement on a 3D in vitro model based on spheroids formed from human induced pluripotent stem cells (hiPSCs). First, we compare self-renewing versus differentiating media conditions in free-floating cultures and observe the emergence of tri-germ layers. In these unconfined conditions, BMP4 exposure induces polarised expression of SOX17 in conjunction with spheroid elongation. We then physically confine spheroids using PEGpeptide hydrogels and observe dramatically reduced SOX17 expression, albeit rescued if gels that soften over time are used instead. Our study combines high-content imaging, synthetic hydrogels, and hiPSCs-derived models of early development to define the drivers that cause changes in the shape and the emergence of germ layers.

    Keywords: Pluripotent Stem Cells, Morphogenesis, germ layer differentiation, PEG-based hydrogels, High content image analysis

    Received: 19 Jul 2024; Accepted: 02 Oct 2024.

    Copyright: © 2024 Alsehli, Roy, Williams, Kuziola, Guo, Green, Gentleman and Danovi. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence:
    Haneen S. Alsehli, Centre for Stem Cell Biology, The University of Sheffield, Sheffield, United Kingdom
    Davide Danovi, Centre for Stem Cells & Regenerative Medicine, School of Basic & Medical Biosciences, Faculty of Life Sciences & Medicine, King's College London, London, WC2R 2LS, England, United Kingdom

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