Improved Characterization of Circulating Tumor Cells and Cancer-Associated Fibroblasts in One-Tube Assay in Breast Cancer Patients Using Imaging Flow Cytometry

Circulating tumor cells (CTCs) and circulating cancer-associated fibroblasts (cCAFs) have been individually considered strong indicators of cancer progression. However, technical limitations prevented their simultaneous analysis in the context CTC phenotypes different from epithelial. This study aimed to analyze CTCs cCAFs simultaneously peripheral blood 210 breast patients using DAPI/pan-keratin (K)/vimentin (V)/alpha-SMA/CD29/CD45/CD31 immunofluorescent staining novel technology—imaging flow cytometry (imFC). Single clustered sizes (i.e., epithelial phenotype K+/V− epithelial–mesenchymal transition (EMT)-related CTCs, such as K+/V+, K−/V+, K−/V−) were detected 27.6% samples correlated with metastases. EMT-related interacted more frequently normal tended occur tumors progressing during therapy, while coincided (mainly seven (3.3%) seemed correlate presence metastases, particularly visceral ones. emphasizes advantages imFC field liquid biopsy highlights importance multimarker-based subpopulations progression-related cells, i.e., cCAFs. The co-detection might improve identification at higher risk progression monitoring therapy.