Assistant Professor Seiichiro Ishihara and Professor Hisashi Haga, participants in Co-Creation Core for Soft Materials Aspiring Research & Translation (C³-SMART), have developed a protocol for culturing cells on collagen gels with adjustable stiffness.

The stiffness of the extracellular matrix (ECM) is extremely important for cellular functions such as cell proliferation and differentiation. In particular, cancer cells are known to change their malignancy in response to ECM stiffness, making it a crucial element in understanding and controlling cellular function.

This study developed a protocol for culturing cells on collagen gels with adjusted stiffness by adding genipin, a low-cytotoxicity amine-based crosslinking agent. This report describes the procedures for preparing the collagen gel using genipin, culturing cells, and immunofluorescence staining of cells on the gel.

This protocol allows for the cultivation of cells on collagen gels of different stiffness with low cytotoxicity. It is expected to have applications in functional analysis and protein/gene expression analysis of cells cultured on extracellular matrices of different stiffness.