Morphogen-driven Differentiation is Precluded by Physical Confinement in Human IPSCs Spheroids

Overview

Journal Front Bioeng Biotechnol

Date 2024 Nov 26

PMID 39588360

Authors

Haneen S Alsehli

Errin Roy

Thomas Williams

Alicja Kuziola

Yunzhe Guo

Cecile A Dreiss

Jeremy B A Green

Eileen Gentleman

Davide Danovi

Affiliations

Soon will be listed here.

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 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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