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Modeling Somitogenesis and Axial Development In Vitro with Axioloids.

Keun-Tae Kim¹, Sofiane Hamidi¹, Cantas Alev²

¹Institute for the Advanced Study of Human Biology (ASHBi), Kyoto University, Kyoto, Japan.

Methods in Molecular Biology (Clifton, N.J.)

|April 22, 2026

Summary

This summary is machine-generated.

Axioloids, 3D stem cell models, mimic human somitogenesis and axial development in vitro. This protocol details their generation for studying developmental disorders.

Keywords:

Axial development Disease modeling Extracellular matrix (ECM)Human pluripotent stem cells (hPSCs)In vitro embryo model Presomitic mesoderm (PSM)Retinoid signaling Segmentation clock Somite epithelialization Somitogenesis

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Area of Science:

Developmental Biology
Stem Cell Biology
Regenerative Medicine

Background:

Human somitogenesis and axial development are complex processes difficult to study in vivo.
Pluripotent stem cells (PSCs) offer a promising avenue for in vitro modeling.
Axioloids are novel 3D structures derived from PSCs that recapitulate key developmental events.

Purpose of the Study:

To provide a detailed protocol for generating axioloids from PSCs.
To establish axioloids as a robust in vitro model for human axial development.
To facilitate the study of congenital disorders affecting the spine and axial skeleton.

Main Methods:

Derivation of axioloids from pluripotent stem cells (PSCs).
Characterization of axioloids for key developmental features: axial elongation, somite formation, rostrocaudal patterning, segmentation clock activity, signaling gradients (FGF/WNT, retinoic acid), and HOX gene expression.
In vitro recapitulation of somitogenesis and early axis development.

Main Results:

Axioloids successfully model human somitogenesis and early axial development in vitro.
These 3D structures exhibit essential morphogenetic features, including epithelial somite formation and rostrocaudal patterning.
Axioloids reproduce in vivo-like spatiotemporal signaling gradients and HOX gene expression patterns.

Conclusions:

Axioloids represent a versatile and reproducible platform for studying human axial development.
This model system is valuable for investigating congenital disorders of the spine and axial skeleton.
The provided protocol enables standardized generation and characterization of axioloids for research purposes.