A synthetic hydrogel scaffold enhancing stem cell signatures in colorectal cancer cells.

Colorectal cancer (CRC) is one of the most prevalent and lethal malignancies worldwide. Strategies to eradicate cancer stem cells (CSCs) are essential for developing more promising therapies against CRC. However, the rarity and heterogeneity of these cells within tumor masses have impeded a deeper understanding of CSCs. Here, we show that cultivation on synthetic hydrogel scaffolds induces reprogramming of conventional CRC cell lines into a CSC-like state. We found that culture on a polyampholyte hydrogel with a net positive charge enhanced the expression of several stem cell markers in CRC cells. Whole transcriptome analysis revealed that signatures related to CSCs and intestinal stem cells were globally upregulated by hydrogel culture. As a result of this transcriptional reprogramming, CRC cells exhibited a metabolic shift from glycolysis to oxidative phosphorylation. Moreover, we identified MEF2D as a key transcription factor that promotes the CSC signature in CRC. Together, these results establish a hydrogel culture system as a versatile platform for inducing CSC traits in CRC cells and demonstrate its usefulness in elucidating mechanisms supporting CSCs.