An innovative in vitro model for studying the biology of cardiac fibroblasts originating from the epicardium
Muller-Sanchez, C.; Muniz-Banciella, M. G.; Reina, M.; Soriano, F. X.; Martinez-Estrada, O. M.
Show abstract
Embryonic epicardium is a major source of cardiac fibroblasts (CFs), which play essential roles in heart development and response to heart injury. In this study, we developed a novel mouse model to identify distinct populations of epicardium-derived CFs. Our Wt1GFP/+;Wt1Cre;ROSA26-tdRFP model enables lineage tracing of WT1Cre-labeled (RFP+) fibroblasts and the identification of cells actively expressing WT1 (GFP+). Flow cytometry at early postnatal stages showed that RFP+ cells form a heterogeneous stromal population, with 20.13% co-expressing GFP, indicating persistent WT1 expression in a subset. We successfully immortalized RFP+ cardiac stromal cells, highly enriched in fibroblasts, by excluding other Wt1Cre-active cell types. Through culture condition optimization, we could selectively expand or differentiate specific fibroblast subpopulations, increasing the models utility. These immortalized cells, carrying an integrated WT1 reporter system, provide a robust in vitro platform to study fibroblast activation, differentiation, and plasticity under defined conditions. Summary blurbThis study presents a novel in vitro mouse model for investigating the activation, identity, and functional properties of epicardium-derived cardiac fibroblasts.
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