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The absence of a universal classification for CAFs hinders their research and therapeutic targeting. To define CAF phenotypes, we developed patient-derived cell cultures of breast cancer (BC) and validated and characterized four distinct CAF subtypes (S1\u2013S4) by Costa\u2019s classification. Three out of five primary cell cultures of BC demonstrated different functional features rather than fixed cellular states due to the plasticity of the CAF phenotype. CAF crosstalk with cancer cells supported their survival in the presence of anticancer drugs. Based on the analysis of the cytotoxic effect of doxorubicin, cisplatin and tamoxifen, it was demonstrated that CAF-S4 and CAF-S1 cells were sensitive to the action of all drugs investigated, despite the fact that they possessed different mechanisms of action. CAF-S2 cells exhibited the highest level of resistance to the antitumour agents. Homotypic and heterotypic spheroids with CAFs could be used to model the fibrotic area of BC in vitro. The patient-derived cell cultures of CAFs formed spheroids. Hypoxia-activated CAF-S4 have been shown to stimulate the metastatic potential of triple-negative BC cells in a heterotypic spheroid model. Consequently, this study could be a starting point for the development of novel therapeutic strategies that target CAFs and their interactions with cancer cells.<\/jats:p>","DOI":"10.3390\/ijms26167789","type":"journal-article","created":{"date-parts":[[2025,8,13]],"date-time":"2025-08-13T07:45:31Z","timestamp":1755071131000},"page":"7789","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Exploring the Heterogeneity of Cancer-Associated Fibroblasts via Development of Patient-Derived Cell Culture of Breast Cancer"],"prefix":"10.3390","volume":"26","author":[{"given":"Anna","family":"Ilyina","sequence":"first","affiliation":[{"name":"Scientific Center of Genetics and Life Sciences, Sirius University of Science and Technology, 1 Olimpiysky Avenue, Sirius 354340, Russia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0009-0005-1477-1118","authenticated-orcid":false,"given":"Anastasia","family":"Leonteva","sequence":"additional","affiliation":[{"name":"Scientific Center of Genetics and Life Sciences, Sirius University of Science and Technology, 1 Olimpiysky Avenue, Sirius 354340, Russia"},{"name":"Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of Sciences, Akad. 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Clin."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"374","DOI":"10.1016\/j.ccell.2023.02.016","article-title":"The Evolving Tumor Microenvironment: From Cancer Initiation to Metastatic Outgrowth","volume":"41","author":"Joyce","year":"2023","journal-title":"Cancer Cell"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Zheng, J., and Hao, H. (2024). The Importance of Cancer-Associated Fibroblasts in Targeted Therapies and Drug Resistance in Breast Cancer. Front. Oncol., 13.","DOI":"10.3389\/fonc.2023.1333839"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1158\/2326-6066.CIR-14-0209","article-title":"Tumors: Wounds That Do Not Heal\u2014Redux","volume":"3","author":"Dvorak","year":"2015","journal-title":"Cancer Immunol. Res."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1161","DOI":"10.1016\/j.stem.2022.07.006","article-title":"Wound Healing, Fibroblast Heterogeneity, and Fibrosis","volume":"29","author":"Talbott","year":"2022","journal-title":"Cell Stem Cell"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"624","DOI":"10.1016\/j.omto.2022.02.003","article-title":"Integrated Analysis Reveals the Molecular Features of Fibrosis in Triple-Negative Breast Cancer","volume":"24","author":"Ding","year":"2022","journal-title":"Mol. Ther.-Oncolytics"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"5094","DOI":"10.1158\/1078-0432.CCR-24-1518","article-title":"High Mechanical Conditioning by Tumor Extracellular Matrix Stiffness Is a Predictive Biomarker for Antifibrotic Therapy in HER2-Negative Breast Cancer","volume":"30","author":"Bermejo","year":"2024","journal-title":"Clin. Cancer Res."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Lugo-Cintr\u00f3n, K.M., Gong, M.M., Ayuso, J.M., Tomko, L.A., Beebe, D.J., Virumbrales-Mu\u00f1oz, M., and Ponik, S.M. (2020). Breast Fibroblasts and ECM Components Modulate Breast Cancer Cell Migration through the Secretion of MMPs in a 3D Microfluidic Co-Culture Model. Cancers, 12.","DOI":"10.3390\/cancers12051173"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Jia, H., Chen, X., Zhang, L., and Chen, M. (2025). Cancer Associated Fibroblasts in Cancer Development and Therapy. J. Hematol. Oncol., 18.","DOI":"10.1186\/s13045-025-01688-0"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1758","DOI":"10.1038\/s41416-024-02671-1","article-title":"Cancer-Associated Fibroblasts Expressing Fibroblast Activation Protein and Podoplanin in Non-Small Cell Lung Cancer Predict Poor Clinical Outcome","volume":"130","author":"Mathieson","year":"2024","journal-title":"Br. J. Cancer"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"96","DOI":"10.1186\/s12964-023-01125-0","article-title":"What Is New in Cancer-Associated Fibroblast Biomarkers?","volume":"21","author":"Zhao","year":"2023","journal-title":"Cell Commun. Signal."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1480","DOI":"10.1016\/j.ccell.2024.08.011","article-title":"Classifying Cancer-Associated Fibroblasts\u2014The Good, the Bad, and the Target","volume":"42","author":"Cords","year":"2024","journal-title":"Cancer Cell"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2708","DOI":"10.1111\/cas.14537","article-title":"Heterogeneity of Cancer-associated Fibroblasts: Opportunities for Precision Medicine","volume":"111","author":"Kanzaki","year":"2020","journal-title":"Cancer Sci."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"905","DOI":"10.1016\/j.ccell.2025.03.004","article-title":"Conserved Spatial Subtypes and Cellular Neighborhoods of Cancer-Associated Fibroblasts Revealed by Single-Cell Spatial Multi-Omics","volume":"43","author":"Liu","year":"2025","journal-title":"Cancer Cell"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"463","DOI":"10.1016\/j.ccell.2018.01.011","article-title":"Fibroblast Heterogeneity and Immunosuppressive Environment in Human Breast Cancer","volume":"33","author":"Costa","year":"2018","journal-title":"Cancer Cell"},{"key":"ref_16","doi-asserted-by":"crossref","unstructured":"Kazakova, A.N., Lukina, M.M., Anufrieva, K.S., Bekbaeva, I.V., Ivanova, O.M., Shnaider, P.V., Slonov, A., Arapidi, G.P., and Shender, V.O. (2024). Exploring the Diversity of Cancer-Associated Fibroblasts: Insights into Mechanisms of Drug Resistance. Front. Cell Dev. Biol., 12.","DOI":"10.3389\/fcell.2024.1403122"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2094","DOI":"10.1158\/1078-0432.CCR-21-3570","article-title":"Immunostimulatory Cancer-Associated Fibroblast Subpopulations Can Predict Immunotherapy Response in Head and Neck Cancer","volume":"28","author":"Obradovic","year":"2022","journal-title":"Clin. Cancer Res."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Louault, K., Li, R.-R., and DeClerck, Y.A. (2020). Cancer-Associated Fibroblasts: Understanding Their Heterogeneity. Cancers, 12.","DOI":"10.3390\/cancers12113108"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1971","DOI":"10.1002\/ijc.29939","article-title":"CD90+ Stromal Cells Are the Major Source of IL-6 Which Supports Cancer Stem-like Cells and Inflammation in Colorectal Cancer","volume":"138","author":"Huynh","year":"2016","journal-title":"Int. J. Cancer"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"46","DOI":"10.1186\/s12935-019-0766-5","article-title":"Establishment of Primary Human Breast Cancer Cell Lines Using \u201cPulsed Hypoxia\u201d Method and Development of Metastatic Tumor Model in Immunodeficient Mice","volume":"19","author":"Nushtaeva","year":"2019","journal-title":"Cancer Cell Int."},{"key":"ref_21","doi-asserted-by":"crossref","unstructured":"Nushtaeva, A., Ermakov, M., Abdurakhmanova, M., Troitskaya, O., Belovezhets, T., Varlamov, M., Gayner, T., Richter, V., and Koval, O. (2023). \u201cPulsed Hypoxia\u201d Gradually Reprograms Breast Cancer Fibroblasts into pro-Tumorigenic Cells via Mesenchymal\u2013Epithelial Transition. Int. J. Mol. Sci., 24.","DOI":"10.3390\/ijms24032494"},{"key":"ref_22","first-page":"91","article-title":"The Optimization of Methods for the Establishment of Heterogeneous Three-Dimensional Cellular Models of Breast Cancer","volume":"17","author":"Nushtaeva","year":"2023","journal-title":"Genes Cells"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"199","DOI":"10.3390\/bios5020199","article-title":"Application of xCELLigence RTCA Biosensor Technology for Revealing the Profile and Window of Drug Responsiveness in Real Time","volume":"5","author":"Kho","year":"2015","journal-title":"Biosensors"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"8809","DOI":"10.1039\/D0TB01454K","article-title":"Real-Time Monitoring of Human Schwann Cells on Heparin-Collagen Coatings Reveals Enhanced Adhesion and Growth Factor Response","volume":"8","author":"Almodovar","year":"2020","journal-title":"J. Mater. Chem. B"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"345","DOI":"10.4103\/0973-1482.157301","article-title":"Sensitivity of Endometrial Cancer Cells from Primary Human Tumor Samples to New Potential Anticancer Peptide Lactaptin","volume":"11","author":"Koval","year":"2015","journal-title":"J. Cancer Res. Ther."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"159","DOI":"10.5603\/RPOR.a2023.0026","article-title":"Primary Cancer-Associated Fibroblasts Exhibit High Heterogeneity among Breast Cancer Subtypes","volume":"28","author":"Piwocka","year":"2023","journal-title":"Rep. Pract. Oncol. Radiother."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Ostrowska-Podhorodecka, Z., Ding, I., Norouzi, M., and McCulloch, C.A. (2022). Impact of Vimentin on Regulation of Cell Signaling and Matrix Remodeling. Front. Cell Dev. Biol., 10.","DOI":"10.3389\/fcell.2022.869069"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Jiang, D., and Rinkevich, Y. (2018). Defining Skin Fibroblastic Cell Types beyond CD90. Front. Cell Dev. Biol., 6.","DOI":"10.3389\/fcell.2018.00133"},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Blaschuk, O.W. (2022). Potential Therapeutic Applications of N-Cadherin Antagonists and Agonists. Front. Cell Dev. Biol., 10.","DOI":"10.3389\/fcell.2022.866200"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1244","DOI":"10.1002\/cac2.12495","article-title":"CD146 Promotes Malignant Progression of Breast Phyllodes Tumor through Suppressing DCBLD2 Degradation and Activating the AKT Pathway","volume":"43","author":"Chen","year":"2023","journal-title":"Cancer Commun."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"6073","DOI":"10.1158\/0008-5472.CAN-18-0136","article-title":"HER2\/EGFR\u2013AKT Signaling Switches TGF\u03b2 from Inhibiting Cell Proliferation to Promoting Cell Migration in Breast Cancer","volume":"78","author":"Huang","year":"2018","journal-title":"Cancer Res."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"617","DOI":"10.1038\/s41580-024-00716-0","article-title":"Fibroblast and Myofibroblast Activation in Normal Tissue Repair and Fibrosis","volume":"25","author":"Younesi","year":"2024","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"15593","DOI":"10.18632\/oncotarget.14777","article-title":"Differential Expression of CD44 and CD24 Markers Discriminates the Epitheliod from the Fibroblastoid Subset in a Sarcomatoid Renal Carcinoma Cell Line: Evidence Suggesting the Existence of Cancer Stem Cells in Both Subsets as Studied with Sorted Cells","volume":"8","author":"Hsieh","year":"2017","journal-title":"Oncotarget"},{"key":"ref_34","first-page":"4411","article-title":"Biological Heterogeneity of Primary Cancer-Associated Fibroblasts Determines the Breast Cancer Microenvironment","volume":"12","author":"Musielak","year":"2022","journal-title":"Am. J. Cancer Res."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"66","DOI":"10.14216\/kjco.22009","article-title":"FSP-1 Expression in Cancer Cells Is Relevant to Long-Term Oncological Outcomes in Nonmetastatic Colorectal Cancer","volume":"18","author":"Im","year":"2022","journal-title":"Korean J. Clin. Oncol."},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Takahashi, N., Yokoi, S., Kimura, H., Naiki, H., Matsusaka, T., Yamamoto, Y., Nakatani, K., Kasuno, K., and Iwano, M. (2023). Renoprotective Effects of Extracellular Fibroblast Specific Protein 1 via Nuclear Factor Erythroid 2-Related Factor-Mediated Antioxidant Activity. Sci. Rep., 13.","DOI":"10.1038\/s41598-023-49863-y"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Avagliano, A., Fiume, G., Ruocco, M.R., Martucci, N., Vecchio, E., Insabato, L., Russo, D., Accurso, A., Masone, S., and Montagnani, S. (2020). Influence of Fibroblasts on Mammary Gland Development, Breast Cancer Microenvironment Remodeling, and Cancer Cell Dissemination. Cancers, 12.","DOI":"10.3390\/cancers12061697"},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Hillsley, A., Santos, J.E., and Rosales, A.M. (2021). A Deep Learning Approach to Identify and Segment Alpha-Smooth Muscle Actin Stress Fiber Positive Cells. Sci. Rep., 11.","DOI":"10.1038\/s41598-021-01304-4"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1186\/s12935-025-03689-y","article-title":"Doxorubicin Loaded Exosomes Inhibit Cancer-Associated Fibroblasts Growth: In Vitro and in Vivo Study","volume":"25","author":"Attar","year":"2025","journal-title":"Cancer Cell Int."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"555","DOI":"10.1134\/S1990519X22060050","article-title":"Breast Cancer Cells in 3D Model Alters Their Sensitivity to Hormonal and Growth Factors","volume":"16","author":"Nushtaeva","year":"2022","journal-title":"Cell Tiss. Biol."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Harper, J.M. (2024). Primary Cell Culture as a Model System for Evolutionary Molecular Physiology. Int. J. Mol. Sci., 25.","DOI":"10.3390\/ijms25147905"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"23","DOI":"10.14744\/lhhs.2025.37221","article-title":"Patient-Derived Primary Cultures in Breast Cancer: A Comparative Study of Isolation and Culture Methods","volume":"5","year":"2025","journal-title":"Lokman Hekim Health Sci."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"101517","DOI":"10.1016\/j.tice.2021.101517","article-title":"A Not so Worthless Attempt to Develop Primary Culture from Breast FNAC\/CNB Samples!","volume":"71","author":"Goel","year":"2021","journal-title":"Tissue Cell"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"827","DOI":"10.1186\/s12967-023-04714-2","article-title":"Crosstalk and Plasticity Driving between Cancer-Associated Fibroblasts and Tumor Microenvironment: Significance of Breast Cancer Metastasis","volume":"21","author":"Zhang","year":"2023","journal-title":"J. Transl. Med."},{"key":"ref_45","doi-asserted-by":"crossref","unstructured":"Romano, V., Ruocco, M.R., Carotenuto, P., Barbato, A., Venuta, A., Acampora, V., De Lella, S., Vigliar, E., Iaccarino, A., and Troncone, G. (2022). Generation and Characterization of a Tumor Stromal Microenvironment and Analysis of Its Interplay with Breast Cancer Cells: An in Vitro Model to Study Breast Cancer-Associated Fibroblast Inactivation. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23126875"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1391","DOI":"10.1016\/j.jcmgh.2023.02.014","article-title":"Mimicking Tumor Cell Heterogeneity of Colorectal Cancer in a Patient-Derived Organoid-Fibroblast Model","volume":"15","author":"Atanasova","year":"2023","journal-title":"Cell. Mol. Gastroenterol. Hepatol."},{"key":"ref_47","doi-asserted-by":"crossref","unstructured":"Cao, Z., Quazi, S., Arora, S., Osellame, L.D., Burvenich, I.J., Janes, P.W., and Scott, A.M. (2025). Cancer-Associated Fibroblasts as Therapeutic Targets for Cancer: Advances, Challenges, and Future Prospects. J. Biomed. Sci., 32.","DOI":"10.1186\/s12929-024-01099-2"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"148","DOI":"10.1038\/s41392-020-00259-8","article-title":"CD146, from a Melanoma Cell Adhesion Molecule to a Signaling Receptor","volume":"5","author":"Wang","year":"2020","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_49","doi-asserted-by":"crossref","unstructured":"Lan, X., Li, W., Zhao, K., Wang, J., Li, S., and Zhao, H. (2025). Revisiting the Role of Cancer-Associated Fibroblasts in Tumor Microenvironment. Front. Immunol., 16.","DOI":"10.3389\/fimmu.2025.1582532"},{"key":"ref_50","doi-asserted-by":"crossref","unstructured":"Kang, M., Min, C., Somayadineshraj, D., and Shin, J.H. (2024). AI-Driven Classification of Cancer-Associated Fibroblasts Using Morphodynamic and Motile Features. bioRxiv.","DOI":"10.1101\/2024.02.22.581611"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"559","DOI":"10.1038\/s41597-024-03399-6","article-title":"Direct Comparison of Mass Cytometry and Single-Cell RNA Sequencing of Human Peripheral Blood Mononuclear Cells","volume":"11","author":"Su","year":"2024","journal-title":"Sci. Data"},{"key":"ref_52","doi-asserted-by":"crossref","unstructured":"Yang, J., Xin, B., Wang, X., and Wan, Y. (2025). Cancer-Associated Fibroblasts in Breast Cancer in the Single-Cell Era: Opportunities and Challenges. Biochim. Biophys. Acta (BBA)-Rev. Cancer, 1880.","DOI":"10.1016\/j.bbcan.2025.189291"},{"key":"ref_53","doi-asserted-by":"crossref","unstructured":"Xia, Z., and De Wever, O. (2025). The Plasticity of Cancer-Associated Fibroblasts. Trends Cancer.","DOI":"10.1016\/j.trecan.2025.04.012"},{"key":"ref_54","doi-asserted-by":"crossref","unstructured":"Hogstrom, J., Cruz, K., Patel, J., Mehta, T., Warren, A., and Muranen, T. (2022). Abstract 3462: Modeling Drug Resistance in Hormone Receptor Positive Breast Cancer Using Patient Derived Organoid Cultures and Cancer Associated Fibroblasts. Cancer Res., 82.","DOI":"10.1158\/1538-7445.AM2022-3462"},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Zhao, Z., Li, T., Sun, L., Yuan, Y., and Zhu, Y. (2023). Potential Mechanisms of Cancer-Associated Fibroblasts in Therapeutic Resistance. Biomed. Pharmacother., 166.","DOI":"10.1016\/j.biopha.2023.115425"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"28","DOI":"10.1186\/s12935-023-03190-4","article-title":"Navigating Challenges: Optimising Methods for Primary Cell Culture Isolation","volume":"24","author":"Piwocka","year":"2024","journal-title":"Cancer Cell Int."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"16","DOI":"10.1172\/JCI93554","article-title":"The Fibrotic Tumor Stroma","volume":"128","author":"Yamauchi","year":"2018","journal-title":"J. Clin. Investig."},{"key":"ref_58","doi-asserted-by":"crossref","unstructured":"Barbosa, M.A.G., Xavier, C.P.R., Pereira, R.F., Petrikait\u0117, V., and Vasconcelos, M.H. (2021). 3D Cell Culture Models as Recapitulators of the Tumor Microenvironment for the Screening of Anti-Cancer Drugs. Cancers, 14.","DOI":"10.3390\/cancers14010190"},{"key":"ref_59","doi-asserted-by":"crossref","unstructured":"Pulak, R., Shah, S., Kim, Y., and D\u2019Souza, G.G. (2025). Development of Triple Co-Culture Spheroid Models of Lung and Ovarian Carcinomas for in Vitro Evaluation of Antitumor Therapies. Cancer Res., 85.","DOI":"10.1158\/1538-7445.AM2025-6465"},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"2402946","DOI":"10.1002\/adhm.202402946","article-title":"Immune-infiltrated Cancer Spheroid Model with Vascular Recirculation Reveals Temporally Dependent and Tissue-specific Macrophage Recruitment","volume":"14","author":"Zhang","year":"2025","journal-title":"Adv. Healthc. Mater."},{"key":"ref_61","doi-asserted-by":"crossref","unstructured":"Nishikiori, N., Takada, K., Sato, T., Miyamoto, S., Watanabe, M., Hirakawa, Y., Sekiguchi, S., Furuhashi, M., Yorozu, A., and Takano, K. (2023). Physical Properties and Cellular Metabolic Characteristics of 3D Spheroids Are Possible Definitive Indices for the Biological Nature of Cancer-Associated Fibroblasts. Cells, 12.","DOI":"10.3390\/cells12172160"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"e33585","DOI":"10.1016\/j.heliyon.2024.e33585","article-title":"Lung Decellularized Matrix-Derived 3D Spheroids: Exploring Silicosis through the Impact of the Nrf2\/Bax Pathway on Myofibroblast Dynamics","volume":"10","author":"Xue","year":"2024","journal-title":"Heliyon"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"3444","DOI":"10.1016\/j.yexcr.2008.09.004","article-title":"Formation and Activation of Fibroblast Spheroids Depend on Fibronectin\u2013Integrin Interaction","volume":"314","author":"Kankuri","year":"2008","journal-title":"Exp. Cell Res."},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"17038","DOI":"10.1038\/cddiscovery.2017.38","article-title":"Generation and Analysis of Spheroids from Human Primary Skin Myofibroblasts: An Experimental System to Study Myofibroblasts Deactivation","volume":"3","author":"Granato","year":"2017","journal-title":"Cell Death Discov."},{"key":"ref_65","doi-asserted-by":"crossref","unstructured":"Du, W., Novin, A., Liu, Y., Afzal, J., Liu, S., Suhail, Y. (2024). Stable and Oscillatory Hypoxia Differentially Regulate Invasibility of Breast Cancer-Associated Fibroblasts. Mechanobiol. Med., 2.","DOI":"10.1101\/2024.03.26.586706"},{"key":"ref_66","doi-asserted-by":"crossref","unstructured":"Mehta, P., Bordoloi, A.D., Ravensbergen, C., David, M.K.H., Mesker, W., Liefers, G.J., Ten Dijke, P., and Boukany, P.E. (2025). Inter-Spheroid Proximity and Matrix Remodeling Determine CAF-Mediated Cancer Cell Invasion. bioRxiv.","DOI":"10.1101\/2025.04.24.648914"},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"e130751","DOI":"10.1172\/jci.insight.130751","article-title":"Fibroblast Subtypes Define a Metastatic Matrisome in Breast Cancer","volume":"5","author":"Brechbuhl","year":"2020","journal-title":"JCI Insight"},{"key":"ref_68","doi-asserted-by":"crossref","unstructured":"Bakleh, M.Z., and Al Haj Zen, A. (2025). The Distinct Role of HIF-1\u03b1 and HIF-2\u03b1 in Hypoxia and Angiogenesis. Cells, 14.","DOI":"10.3390\/cells14090673"},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"3432","DOI":"10.3892\/mmr.2015.3815","article-title":"Role of Hypoxia-Inducible Factor-1\u03b1 and CD146 in Epidermal Growth Factor Receptor-Mediated Angiogenesis in Salivary Gland Adenoid Cystic Carcinoma","volume":"12","author":"Wang","year":"2015","journal-title":"Mol. Med. Rep."},{"key":"ref_70","doi-asserted-by":"crossref","unstructured":"Dwivedi, N., Bahaar, H., Shashank, D., Cherry, C.E., Gangadharan, C., Suresh, A., Kuriakose, M.A., Pillai, V., Das, M., and Smitha, P.K. (2023). Establishment and Characterization of Three Novel CAF Cell Lines from HNSCC Patients. bioRxiv.","DOI":"10.1101\/2023.01.08.523131"},{"key":"ref_71","doi-asserted-by":"crossref","first-page":"174","DOI":"10.1038\/s41568-019-0238-1","article-title":"A Framework for Advancing Our Understanding of Cancer-Associated Fibroblasts","volume":"20","author":"Sahai","year":"2020","journal-title":"Nat. Rev. Cancer"},{"key":"ref_72","doi-asserted-by":"crossref","first-page":"1866","DOI":"10.3892\/etm.2017.4781","article-title":"iCELLigence Real-Time Cell Analysis System for Examining the Cytotoxicity of Drugs to Cancer Cell Lines","volume":"14","author":"Albeniz","year":"2017","journal-title":"Exp. Ther. Med."},{"key":"ref_73","doi-asserted-by":"crossref","first-page":"10","DOI":"10.1016\/j.mrrev.2012.08.001","article-title":"In Vitro Cell Migration and Invasion Assays","volume":"752","author":"Kramer","year":"2013","journal-title":"Mutat. Res.\/Rev. Mutat. Res."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/16\/7789\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,9]],"date-time":"2025-10-09T18:25:44Z","timestamp":1760034344000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/26\/16\/7789"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2025,8,12]]},"references-count":73,"journal-issue":{"issue":"16","published-online":{"date-parts":[[2025,8]]}},"alternative-id":["ijms26167789"],"URL":"https:\/\/doi.org\/10.3390\/ijms26167789","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2025,8,12]]}}}