Fig. 2

Methodology for studying CAFs. (A) CAFs may be obtained through primary culture, or transformation from other cells, such as normal fibroblasts or pancreatic stellate cells (PSCs), for further researches. (B) Animal models of CAFs include patient-derived xenografts (PDX) and transplanted tumor models established from cell lines. Genetically engineered mouse models (GEMMs) can spontaneously develop tumors, better simulating the interaction between CAFs and TME. These genetically engineered mice also allow for the specific elimination of CAF subsets using drugs such as diphtheria toxin, as well as lineage tracing of CAF subsets through the expression of specific markers. (C) In vitro approaches to study CAF-TME interactions include 2D/3D co-culture, conditioned media transfer, and tumor organoid models. In vivo methods involve co-inoculation of CAFs and tumor cells. (D) CAF research integrates techniques such as western blotting (WB), flow cytometry, and immunofluorescence. Advanced tools like single-cell RNA sequencing (scRNA-seq) and spatial transcriptomics provide deeper functional insights. This figure was created with BioRender.com