PEER-REVIEWED PUBLICATION

2023

TGF-β2 Induces Epithelial–Mesenchymal Transitions in 2D Planar and 3D Spheroids of the Human Corneal Stroma Fibroblasts in Different Manners

Umetsu A, Ida Y, et al.

Biomedicines

Sapporo Medical University

RESEARCH SUMMARY

This study investigates how Transforming Growth Factor-β2 (TGF-β2) differentially induces epithelial–mesenchymal transition (EMT) in human corneal stromal fibroblasts (HCSFs) cultured in 2D monolayers versus 3D spheroids. TGF-β2 exposure caused increased extracellular matrix (ECM) deposition, actin remodeling, and elevated mitochondrial and glycolytic activity in 2D HCSFs. In contrast, in 3D spheroids, TGF-β2 led to downsizing, stiffening, and selective modulation of ECM-related genes (COL1, COL6, MMP14, TIMP2), with strong induction of ER stress markers. These findings highlight the distinct biomechanical and transcriptional responses of corneal fibroblasts in planar versus spheroidal environments, advancing understanding of stromal fibrosis and wound healing mechanisms.

A CellScale MicroSquisher was used to quantify the mechanical stiffness of live 3D human corneal stromal fibroblast spheroids following TGF-β2 treatment. Individual spheroids were compressed to 50% of their original diameter while measuring force–displacement responses in the ultra-low force regime. These CellScale measurements revealed concentration-dependent spheroid stiffening induced by TGF-β2, providing direct mechanical evidence of EMT-associated tissue remodeling. The MicroSquisher data were essential for demonstrating that mechanical stiffening in 3D spheroids occurs independently of the pronounced ECM gene upregulation observed in 2D cultures, reinforcing the importance of 3D biomechanical context in corneal fibrosis research.

AUTHORS

Araya Umetsu, Yosuke Ida, Tatsuya Sato, Masato Furuhashi, Hiroshi Ohguro, Megumi Watanabe.

PUBLICATION DETAILS

JOURNAL

Biomedicines

YEAR

2023

INSTITUTIONS

Sapporo Medical University

COUNTRIES

Japan

INSTRUMENT USED

MicroSquisher

TESTING METHODS

Compression TestingMicro-Mechanical TestingUltra Low Force Testing

RESEARCH APPLICATIONS

Fibrosis & Tissue RemodelingMechanotransductionMicrotissue and Spheroid MechanicsOphthalmic Biomechanics & Corneal Tissue Engineering

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