Single-Cell Analysis Reveals the Impact of tumor-derived matrix on Cancer-Associated Fibroblasts

High-grade serous ovarian cancer (HGSOC) is a leading cause of death among gynecological malignancies, primarily due to its frequent metastasis to the omentum. To understand its underlying biological processes, in vitro models that can recapitulate the original microenvironment of this organ are essential. Thus, Omentum-derived Gel (OmGel), a matrix derived from tumour-associated omental tissue of HGSOC patients has been developed, which has unprecedented similarity to the in vivo extracellular matrix (ECM). This study explores the responses of cancer-associated fibroblasts (CAFs) on OmGel in comparison to Matrigel, a commonly used matrix for cell culture. We employed single-cell Raman spectroscopy to analyze the biochemical profiles of CAFs cultured on both matrices, categorizing cells into Long, Round, and Extend morphologies based on form factors. Our findings reveal that OmGel induces significant biochemical changes in CAFs, particularly in elongated cells, which exhibit elevated levels of DNA/RNA and lipids compared to those on Matrigel. These results underscore the importance of using physiologically relevant matrices like OmGel for better understanding of cell and matrix interactions in the tumor microenvironment, offering insights into the development of more effective therapeutic strategies targeting the tumor stroma.