biostudies-literatureUnknown32Liao CYIn fibrotic tissues, activated fibroblasts remodel the collagen-rich extracellular matrix (ECM). Intervening with this process represents a candidate therapeutic strategy to attenuate disease progression. Models that generate quantitative data on 3D fibroblast-mediated ECM remodeling with the reproducibility and throughput needed for drug testing are lacking. Here, we develop a model that fits this purpose and produces combined quantitative information on drug efficacy and cytotoxicity. We use microinjection robotics to design patterns of fibrillar collagen-embedded fibroblast clusters and apply automated microscopy and image analysis to quantify ECM remodeling between-, and cell viability within clusters of TGFβ-activated primary human skin or lung fibroblasts. We apply this assay to compMaterials today. Bio101800https://www.ebi.ac.uk/biostudies/studies/S-EPMC12059351biostudies-literatureNovel high throughput 3D ECM remodeling assay identifies MEK as key driver of fibrotic fibroblast activity.PMC12059351Ten Dijke PCrawford JHundscheid JHMDanen EHJCoornaert BLiao CYfalseNovel high throughput 3D ECM remodeling assay identifies MEK as key driver of fibrotic fibroblast activity.In fibrotic tissues, activated fibroblasts remodel the collagen-rich extracellular matrix (ECM). Intervening with this process represents a candidate therapeutic strategy to attenuate disease progression. Models that generate quantitative data on 3D fibroblast-mediated ECM remodeling with the reproducibility and throughput needed for drug testing are lacking. Here, we develop a model that fits this purpose and produces combined quantitative information on drug efficacy and cytotoxicity. We use microinjection robotics to design patterns of fibrillar collagen-embedded fibroblast clusters and apply automated microscopy and image analysis to quantify ECM remodeling between-, and cell viability within clusters of TGFβ-activated primary human skin or lung fibroblasts. We apply this assay to comp2025-01-01T00:00:00Z2025 Jun2026-06-01T10:24:39.046Z2026-04-08T11:20:05.069ZS-EPMC120593514034316410.1016/j.mtbio.2025.101800