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Stiffness-Modulation of Collagen Gels by Genipin-Crosslinking for Cell Culture

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Title: Stiffness-Modulation of Collagen Gels by Genipin-Crosslinking for Cell Culture
Authors: Ishihara, Seiichiro Browse this author →KAKEN DB
Kurosawa, Haruna Browse this author
Haga, Hisashi Browse this author →KAKEN DB
Keywords: collagen gel
extracellular matrix
cell culture
cell morphology
mesenchymal stromal cells
cancer cells
Issue Date: Feb-2023
Publisher: MDPI
Journal Title: Gels
Volume: 9
Issue: 2
Start Page: 148
Publisher DOI: 10.3390/gels9020148
Abstract: The stiffness of extracellular matrices (ECMs) is critical for cellular functions. Therefore, modulating the stiffness of ECMs in vitro is necessary to investigate the role of stiffness in cellular phenomena. Collagen gels are widely used for cell culture matrices in vitro. However, modulation of the stiffness in collagen gels for cell culture is challenging owing to the limited knowledge of the method to increase the stiffness while maintaining low cytotoxicity. Here, we established a novel method to modulate collagen gel stiffness from 0.0292 to 12.5 kPa with low cytotoxicity. We prepared collagens with genipin, a low-cytotoxic crosslinker of amines, at different concentrations and successfully modulated the stiffness of the gels. In addition, on 10 mM genipin-mixed collagen gels (approximately 12.5 kPa), H1299 human lung cancer cells showed spreading morphology and nuclear localization of yes-associated protein (YAP), typical phenomena of cells on stiff ECMs. Mouse mesenchymal stromal cells on 10 mM genipin-mixed collagen gels differentiated to vascular smooth muscle cells. On the other hand, the cells on 0 mM genipin-mixed collagen gels (approximately 0.0292 kPa) differentiated to visceral smooth muscle cells. Our new method provides a novel way to prepare stiffness-modulated collagen gels with low cytotoxicity in cell culture.
Type: article
Appears in Collections:生命科学院・先端生命科学研究院 (Graduate School of Life Science / Faculty of Advanced Life Science) > 雑誌発表論文等 (Peer-reviewed Journal Articles, etc)

Submitter: 石原 誠一郎

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