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

Eileen Gentleman; Davide Danovi; Haneen Alsehli; Alicja Kuziola; Yunzhe Guo; Cecile Dreiss; Jeremy Green; Errin Roy; Tom Williams

doi:10.3389/fbioe.2024.1467412

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

Eileen Gentleman, Davide Danovi^*, Haneen Alsehli, Alicja Kuziola, Yunzhe Guo, Cecile Dreiss, Jeremy Green, Errin Roy, Tom Williams

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract

Introduction: 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. Methods: 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). Results: 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 PEG-peptide hydrogels and observe dramatically reduced SOX17 expression, albeit rescued if gels that soften over time are used instead. Discussion: 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.

Original language	English
Article number	1467412
Journal	Frontiers in Bioengineering and Biotechnology
Volume	12
DOIs	https://doi.org/10.3389/fbioe.2024.1467412
Publication status	Published - 11 Nov 2024

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title = "Morphogen-driven differentiation is precluded by physical confinement in human iPSCs spheroids",

abstract = "Introduction: 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. Methods: 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). Results: 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 PEG-peptide hydrogels and observe dramatically reduced SOX17 expression, albeit rescued if gels that soften over time are used instead. Discussion: 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.",

author = "Eileen Gentleman and Davide Danovi and Haneen Alsehli and Alicja Kuziola and Yunzhe Guo and Cecile Dreiss and Jeremy Green and Errin Roy and Tom Williams",

note = "Publisher Copyright: Copyright {\textcopyright} 2024 Alsehli, Roy, Williams, Kuziola, Guo, Dreiss, Green, Gentleman and Danovi.",

year = "2024",

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doi = "10.3389/fbioe.2024.1467412",

language = "English",

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T1 - Morphogen-driven differentiation is precluded by physical confinement in human iPSCs spheroids

AU - Gentleman, Eileen

AU - Danovi, Davide

AU - Alsehli, Haneen

AU - Kuziola, Alicja

AU - Guo, Yunzhe

AU - Dreiss, Cecile

AU - Green, Jeremy

AU - Roy, Errin

AU - Williams, Tom

PY - 2024/11/11

Y1 - 2024/11/11

N2 - Introduction: 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. Methods: 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). Results: 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 PEG-peptide hydrogels and observe dramatically reduced SOX17 expression, albeit rescued if gels that soften over time are used instead. Discussion: 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.

AB - Introduction: 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. Methods: 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). Results: 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 PEG-peptide hydrogels and observe dramatically reduced SOX17 expression, albeit rescued if gels that soften over time are used instead. Discussion: 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.

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DO - 10.3389/fbioe.2024.1467412

M3 - Article

SN - 2296-4185

VL - 12

JO - Frontiers in Bioengineering and Biotechnology

JF - Frontiers in Bioengineering and Biotechnology

M1 - 1467412

ER -

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

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