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While traditional studies mostly use 2D cell cultures, 3D models better mimic in vivo tumor conditions. In this study, we generated and characterized 3D spheroids from breast cancer cell lines representing different molecular subtypes. Morphologically, spheroids were either compact (MCF-7\/AZ, T47D, BT474, MDA-IBC-3, BT-20, SUM149PT) or loosely adhered (MDA-MB-468, SK-BR-3, MDA-MB-231), while retaining key parental subtype biomarkers. Cell viability decreased with increasing spheroid size, but apoptotic cCasp3 staining was restricted to Basal-like spheroids. Compact spheroids expressed E- and\/or P-cadherin, indicating epithelial or epithelial\u2013mesenchymal transition (EMT) hybrid traits, while loose spheroids showed vimentin expression linked to a mesenchymal phenotype. In conclusion, EMT-associated features, rather than intrinsic molecular subtype, may contribute to 3D spheroid architecture of breast cancer cell lines.<\/jats:p>","DOI":"10.3390\/ijms27083529","type":"journal-article","created":{"date-parts":[[2026,4,15]],"date-time":"2026-04-15T14:02:33Z","timestamp":1776261753000},"page":"3529","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["From Luminal to Triple Negative: 3D Spheroids Reveal Molecular and Phenotypic Differences Across Breast Cancer Subtypes"],"prefix":"10.3390","volume":"27","author":[{"given":"Maria Miguel","family":"Castro","sequence":"first","affiliation":[{"name":"Cancer Metastasis, i3S, Institute for Research and Innovation in Health, University of Porto, 4200-135 Porto, Portugal"},{"name":"Faculty of Medicine, University of Porto, 4200-319 Porto, Portugal"},{"name":"Cancer Biology and Epigenetics Group, Research Center of IPO Porto (CI-IPOP)\/CI-IPOP@RISE Health Research Network, Portuguese Oncology Institute of Porto (IPO Porto), 4200-072 Porto, Portugal"},{"name":"Porto Comprehensive Cancer Center Raquel Seruca (P.CCC Raquel Seruca), 4200-072 Porto, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Let\u00edcia","family":"Maretti","sequence":"additional","affiliation":[{"name":"Cancer Metastasis, i3S, Institute for Research and Innovation in Health, University of Porto, 4200-135 Porto, Portugal"},{"name":"Porto Comprehensive Cancer Center Raquel Seruca (P.CCC Raquel Seruca), 4200-072 Porto, Portugal"},{"name":"ICBAS, School of Medicine and Biomedical Sciences, 4050-313 Porto, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3832-2027","authenticated-orcid":false,"given":"Catarina","family":"Esqu\u00edvel","sequence":"additional","affiliation":[{"name":"Cancer Metastasis, i3S, Institute for Research and Innovation in Health, University of Porto, 4200-135 Porto, Portugal"},{"name":"Porto Comprehensive Cancer Center Raquel Seruca (P.CCC Raquel Seruca), 4200-072 Porto, Portugal"},{"name":"ICBAS, School of Medicine and Biomedical Sciences, 4050-313 Porto, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"B\u00e1rbara","family":"Sousa","sequence":"additional","affiliation":[{"name":"Cancer Metastasis, i3S, Institute for Research and Innovation in Health, University of Porto, 4200-135 Porto, Portugal"},{"name":"Porto Comprehensive Cancer Center Raquel Seruca (P.CCC Raquel Seruca), 4200-072 Porto, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4186-5345","authenticated-orcid":false,"given":"Carmen","family":"Jer\u00f3nimo","sequence":"additional","affiliation":[{"name":"Cancer Biology and Epigenetics Group, Research Center of IPO Porto (CI-IPOP)\/CI-IPOP@RISE Health Research Network, Portuguese Oncology Institute of Porto (IPO Porto), 4200-072 Porto, Portugal"},{"name":"Porto Comprehensive Cancer Center Raquel Seruca (P.CCC Raquel Seruca), 4200-072 Porto, Portugal"},{"name":"ICBAS, School of Medicine and Biomedical Sciences, 4050-313 Porto, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andrew J.","family":"Ewald","sequence":"additional","affiliation":[{"name":"Department of Cell Biology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA"},{"name":"Giovanis Institute for Translational Cell Biology, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA"},{"name":"Cancer Invasion and Metastasis Program, Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University, Baltimore, MD 21231, USA"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1076-1343","authenticated-orcid":false,"given":"Joana","family":"Paredes","sequence":"additional","affiliation":[{"name":"Cancer Metastasis, i3S, Institute for Research and Innovation in Health, University of Porto, 4200-135 Porto, Portugal"},{"name":"Faculty of Medicine, University of Porto, 4200-319 Porto, Portugal"},{"name":"Porto Comprehensive Cancer Center Raquel Seruca (P.CCC Raquel Seruca), 4200-072 Porto, Portugal"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2026,4,15]]},"reference":[{"key":"ref_1","first-page":"229","article-title":"Global cancer statistics 2022: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries","volume":"74","author":"Bray","year":"2024","journal-title":"CA Cancer J. Clin."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1038\/s41392-024-02108-4","article-title":"Breast cancer: Pathogenesis and treatments","volume":"10","author":"Xiong","year":"2025","journal-title":"Signal Transduct. Target. Ther."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"747","DOI":"10.1038\/35021093","article-title":"Molecular portraits of human breast tumours","volume":"406","author":"Perou","year":"2000","journal-title":"Nature"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"10869","DOI":"10.1073\/pnas.191367098","article-title":"Gene expression patterns of breast carcinomas distinguish tumor subclasses with clinical implications","volume":"98","author":"Sorlie","year":"2001","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"5","DOI":"10.1016\/j.molonc.2010.11.003","article-title":"Deconstructing the molecular portraits of breast cancer","volume":"5","author":"Prat","year":"2011","journal-title":"Mol. Oncol."},{"key":"ref_6","doi-asserted-by":"crossref","unstructured":"Carvalho, E., Canberk, S., Schmitt, F., and Vale, N. (2025). Molecular Subtypes and Mechanisms of Breast Cancer: Precision Medicine Approaches for Targeted Therapies. Cancers, 17.","DOI":"10.3390\/cancers17071102"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"163","DOI":"10.1007\/s10549-023-07094-9","article-title":"Correlation between breast cancer subtypes determined by immunohistochemistry and n-COUNTER PAM50 assay: A real-world study","volume":"203","author":"Jerez","year":"2024","journal-title":"Breast Cancer Res. Treat."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"55","DOI":"10.5858\/2010-0454-RAR.1","article-title":"HER2: Biology, Detection, and Clinical Implications","volume":"135","author":"Gutierrez","year":"2011","journal-title":"Arch. Pathol. Lab. Med."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"339","DOI":"10.1016\/j.breast.2021.07.019","article-title":"Dissecting the biological heterogeneity of HER2-positive breast cancer","volume":"59","author":"Schettini","year":"2021","journal-title":"Breast"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1673","DOI":"10.1056\/NEJMoa052122","article-title":"Trastuzumab plus adjuvant chemotherapy for operable HER2-positive breast cancer","volume":"353","author":"Romond","year":"2005","journal-title":"N. Engl. J. Med."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1659","DOI":"10.1056\/NEJMoa052306","article-title":"Trastuzumab after adjuvant chemotherapy in HER2-positive breast cancer","volume":"353","author":"Procter","year":"2005","journal-title":"N. Engl. J. Med."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"209","DOI":"10.1016\/j.molonc.2010.04.006","article-title":"Triple-negative breast cancer: Present challenges and new perspectives","volume":"4","author":"Podo","year":"2010","journal-title":"Mol. Oncol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"5367","DOI":"10.1158\/1078-0432.CCR-04-0220","article-title":"Immunohistochemical and clinical characterization of the basal-like subtype of invasive breast carcinoma","volume":"10","author":"Nielsen","year":"2004","journal-title":"Clin. Cancer Res."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"R68","DOI":"10.1186\/bcr2635","article-title":"Phenotypic and molecular characterization of the claudin-low intrinsic subtype of breast cancer","volume":"12","author":"Prat","year":"2010","journal-title":"Breast Cancer Res."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1186\/s13045-020-00931-0","article-title":"Modeling neoplastic disease with spheroids and organoids","volume":"13","author":"Zanoni","year":"2020","journal-title":"J. Hematol. Oncol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"306","DOI":"10.1016\/j.addr.2021.03.018","article-title":"The past, present, and future of breast cancer models for nanomedicine development","volume":"173","author":"Vicent","year":"2021","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Katt, M.E., Placone, A.L., Wong, A.D., Xu, Z.S., and Searson, P.C. (2016). In vitro tumor models: Advantages, disadvantages, variables, and selecting the right platform. Front. Bioeng. Biotechnol., 4.","DOI":"10.3389\/fbioe.2016.00012"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"152","DOI":"10.1186\/s12935-021-01853-8","article-title":"Challenges of applying multicellular tumor spheroids in preclinical phase","volume":"21","author":"Han","year":"2021","journal-title":"Cancer Cell Int."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"108","DOI":"10.1016\/j.tibtech.2012.12.003","article-title":"Spheroid culture as a tool for creating 3D complex tissues","volume":"31","author":"Fennema","year":"2013","journal-title":"Trends Biotechnol."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Pinto, B., Henriques, A.C., Silva, P.M.A., and Bousbaa, H. (2020). Three-Dimensional Spheroids as In Vitro Preclinical Models for Cancer Research. Pharmaceutics, 12.","DOI":"10.3390\/pharmaceutics12121186"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/j.jbiotec.2010.01.012","article-title":"Multicellular tumor spheroids: An underestimated tool is catching up again","volume":"148","author":"Hirschhaeuser","year":"2010","journal-title":"J. Biotechnol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1016\/j.ccr.2006.10.008","article-title":"A collection of breast cancer cell lines for the study of functionally distinct cancer subtypes","volume":"10","author":"Neve","year":"2006","journal-title":"Cancer Cell"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"237","DOI":"10.1007\/s10549-013-2743-3","article-title":"Characterization of cell lines derived from breast cancers and normal mammary tissues for the study of the intrinsic molecular subtypes","volume":"142","author":"Prat","year":"2013","journal-title":"Breast Cancer Res. Treat."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"398","DOI":"10.1016\/j.jconrel.2020.04.025","article-title":"Colorectal cancer triple co-culture spheroid model to assess the biocompatibility and anticancer properties of polymeric nanoparticles","volume":"323","author":"Shahbazi","year":"2020","journal-title":"J. Control. Release"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"106560","DOI":"10.1016\/j.ejps.2023.106560","article-title":"Mimicking 3D breast tumor-stromal interactions to screen novel cancer therapeutics","volume":"190","author":"Domingues","year":"2023","journal-title":"Eur. J. Pharm. Sci."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"212","DOI":"10.1016\/j.tcb.2018.12.001","article-title":"EMT Transition States during Tumor Progression and Metastasis","volume":"29","author":"Pastushenko","year":"2019","journal-title":"Trends Cell Biol."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1038\/s41568-021-00332-6","article-title":"Linking EMT programmes to normal and neoplastic epithelial stem cells","volume":"21","author":"Lambert","year":"2021","journal-title":"Nat. Rev. Cancer"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Ribeiro, A.S., and Paredes, J. (2015). P-Cadherin Linking Breast Cancer Stem Cells and Invasion: A Promising Marker to Identify an \u201cIntermediate\/Metastable\u201d EMT State. Front. Oncol., 4.","DOI":"10.3389\/fonc.2014.00371"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1038\/s41523-017-0023-9","article-title":"Inflammatory breast cancer: A model for investigating cluster-based dissemination","volume":"3","author":"Jolly","year":"2017","journal-title":"npj Breast Cancer"},{"key":"ref_30","first-page":"309","article-title":"P-cadherin overexpression is associated with early transformation of the Fallopian tube epithelium and aggressiveness of tubo-ovarian high-grade serous carcinoma","volume":"488","author":"Ribeiro","year":"2025","journal-title":"Virchows Arch."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"27067","DOI":"10.18632\/oncotarget.8166","article-title":"Stability of the hybrid epithelial\/mesenchymal phenotype","volume":"7","author":"Jolly","year":"2016","journal-title":"Oncotarget"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"703","DOI":"10.1002\/cam4.630","article-title":"Three-dimensional microtissues essentially contribute to preclinical validations of therapeutic targets in breast cancer","volume":"5","author":"Falkenberg","year":"2016","journal-title":"Cancer Med."},{"key":"ref_33","unstructured":"Thermo Fisher Scientific (2025, October 06). T47D Cell Line Spheroid Generation and Characterization for HT Assays. Available online: https:\/\/www.thermofisher.com\/pt\/en\/home\/references\/protocols\/cell-culture\/3-d-cell-culture-protocol\/t47d-cell-line-spheroid-generation.html."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"84","DOI":"10.1016\/j.molonc.2007.02.004","article-title":"The morphologies of breast cancer cell lines in three-dimensional assays correlate with their profiles of gene expression","volume":"1","author":"Kenny","year":"2007","journal-title":"Mol. Oncol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1007\/s10911-016-9359-2","article-title":"Generation of Multicellular Breast Cancer Tumor Spheroids: Comparison of Different Protocols","volume":"21","author":"Froehlich","year":"2016","journal-title":"J. Mammary Gland Biol. Neoplasia"},{"key":"ref_36","doi-asserted-by":"crossref","unstructured":"Gong, X., Lin, C., Cheng, J., Su, J., Zhao, H., Liu, T., Wen, X., and Zhao, P. (2015). Generation of Multicellular Tumor Spheroids with Microwell-Based Agarose Scaffolds for Drug Testing. PLoS ONE, 10.","DOI":"10.1371\/journal.pone.0130348"},{"key":"ref_37","doi-asserted-by":"crossref","unstructured":"Cavaco, M., Fraga, P., Valle, J., Andreu, D., Castanho, M.A.R.B., and Neves, V. (2021). Development of breast cancer spheroids to evaluate cytotoxic response to an anticancer peptide. Pharmaceutics, 13.","DOI":"10.3390\/pharmaceutics13111863"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1038\/nmeth1015","article-title":"Three-dimensional culture models of normal and malignant breast epithelial cells","volume":"4","author":"Lee","year":"2007","journal-title":"Nat. Methods"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"134","DOI":"10.21037\/tcr-21-1577","article-title":"Patient-derived tumor spheroid cultures as a promising tool to assist personalized therapeutic decisions in breast cancer","volume":"11","author":"Hofmann","year":"2022","journal-title":"Transl. Cancer Res."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"7259","DOI":"10.1038\/s41598-021-86664-7","article-title":"Differential reprogramming of breast cancer subtypes in 3D cultures and implications for sensitivity to targeted therapy","volume":"11","author":"Koedoot","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1038\/onc.2008.394","article-title":"Comparison of 3D and 2D tumor models reveals enhanced HER2 activation in 3D associated with an increased response to trastuzumab","volume":"28","author":"Pickl","year":"2009","journal-title":"Oncogene"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Krause, S., Maffini, M.V., Soto, A.M., and Sonnenschein, C. (2010). The microenvironment determines the breast cancer cells\u2019 phenotype: Organization of MCF7 cells in 3D cultures. BMC Cancer, 10.","DOI":"10.1186\/1471-2407-10-263"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"1837","DOI":"10.3892\/or.2015.3767","article-title":"Comparison of 2D- and 3D-culture models as drug-testing platforms in breast cancer","volume":"33","author":"Imamura","year":"2015","journal-title":"Oncol. Rep."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"1294","DOI":"10.1038\/s41592-021-01291-4","article-title":"MISpheroID: A knowledgebase and transparency tool for minimum information in spheroid identity","volume":"18","author":"Peirsman","year":"2021","journal-title":"Nat. Methods"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"90","DOI":"10.1016\/j.canlet.2013.10.014","article-title":"T47D breast cancer cells switch from ER\/HER to HER\/c-Src signaling upon acquiring resistance to the antiestrogen fulvestrant","volume":"344","author":"Kirkegaard","year":"2014","journal-title":"Cancer Lett."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"3732","DOI":"10.1158\/0008-5472.CAN-15-2304","article-title":"Three-dimensional breast cancer models mimic hallmarks of size-induced tumor progression","volume":"76","author":"Singh","year":"2016","journal-title":"Cancer Res."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"19950","DOI":"10.1038\/s41598-019-56276-3","article-title":"Up-regulated fibronectin in 3D culture facilitates spreading of triple negative breast cancer cells on 2D through integrin \u03b2-5 and Src","volume":"9","author":"Park","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"412","DOI":"10.5306\/wjco.v5.i3.412","article-title":"Biological subtypes of breast cancer: Prognostic and therapeutic implications","volume":"5","author":"Yersal","year":"2014","journal-title":"World J. Clin. Oncol."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"336","DOI":"10.1039\/C4BM00319E","article-title":"Media additives to promote spheroid circularity and compactness in hanging drop platform","volume":"3","author":"Leung","year":"2014","journal-title":"Biomater. Sci."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"srep19103","DOI":"10.1038\/srep19103","article-title":"3D tumor spheroid models for in vitro therapeutic screening: A systematic approach to enhance the biological relevance of data obtained","volume":"6","author":"Zanoni","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"41237","DOI":"10.1038\/s41598-025-25202-1","article-title":"SpheroidSync as edge cutting transfer strategy for uniform and robust MCF7 spheroids in 3D culture","volume":"15","author":"Mazloomi","year":"2025","journal-title":"Sci. Rep."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"9751","DOI":"10.1038\/s41598-025-92037-1","article-title":"Influence of experimental variables on spheroid attributes","volume":"15","author":"Zhu","year":"2025","journal-title":"Sci. Rep."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"1200","DOI":"10.1038\/modpathol.3800961","article-title":"Immunohistochemical heterogeneity of breast carcinomas negative for estrogen receptors, progesterone receptors and Her2\/neu (basal-like breast carcinomas)","volume":"20","author":"Lerma","year":"2007","journal-title":"Mod. Pathol."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1007\/s10549-009-0545-4","article-title":"Four human breast cancer cell lines with biallelic inactivating \u03b1-catenin gene mutations","volume":"122","author":"Hollestelle","year":"2010","journal-title":"Breast Cancer Res. Treat."},{"key":"ref_55","doi-asserted-by":"crossref","unstructured":"Mangani, S., Koutsakis, C., Koletsis, N.E., Piperigkou, Z., Franchi, M., G\u00f6tte, M., and Karamanos, N.K. (2025). Spheroid-Based 3D Models to Decode Cell Function and Matrix Effectors in Breast Cancer. Cancers, 17.","DOI":"10.3390\/cancers17213512"},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"5395","DOI":"10.2147\/OTT.S249756","article-title":"Characterization of Triple-Negative Breast Cancer MDA-MB-231 Cell Spheroid Model","volume":"13","author":"Huang","year":"2020","journal-title":"OncoTargets Ther."},{"key":"ref_57","doi-asserted-by":"crossref","unstructured":"Arnaldi, P., Zotti, V.D., Bellese, G., Gagliani, M.C., Orecchia, P., Castagnola, P., and Cortese, K. (2025). 3D Breast Cancer Spheroids Reveal Architecture-Dependent HER2 Expression and Signaling. Biology, 14.","DOI":"10.20944\/preprints202510.0712.v1"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"27449","DOI":"10.1038\/s41598-025-12556-9","article-title":"Development of tetraculture spheroids as a versatile 3D model for personalized breast cancer research","volume":"15","author":"Piwocka","year":"2025","journal-title":"Sci. Rep."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"109400","DOI":"10.1016\/j.bej.2024.109400","article-title":"Advanced co-culture 3D breast cancer model to study cell death and nanodrug sensitivity of tumor spheroids","volume":"209","author":"Ahvaraki","year":"2024","journal-title":"Biochem. Eng. J."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"492","DOI":"10.1038\/s41597-025-04832-0","article-title":"Single-cell light-sheet fluorescence 3D images of tumour-stroma spheroid multicultures","volume":"12","author":"Diosdi","year":"2025","journal-title":"Sci. Data"},{"key":"ref_61","doi-asserted-by":"crossref","unstructured":"Saraiva, D.P., Matias, A.T., Braga, S., Jacinto, A., and Cabral, M.G. (2020). Establishment of a 3D Co-culture With MDA-MB-231 Breast Cancer Cell Line and Patient-Derived Immune Cells for Application in the Development of Immunotherapies. Front. Oncol., 10.","DOI":"10.3389\/fonc.2020.01543"},{"key":"ref_62","doi-asserted-by":"crossref","unstructured":"Ortiz, E., Thway, K.H., Ortiz-Soto, G., Yao, P., and Kelber, J.A. (2025). Hetero-multicellular stromal cells incorporate into scaffold-free 3D cultures of epithelial cancer cells to drive invasion. bioRxiv, bioRxiv:2025.01.21.634082.","DOI":"10.1101\/2025.01.21.634082"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"117","DOI":"10.4236\/jct.2022.133009","article-title":"Breast Cancer MCF-7 Cell Spheroid Culture for Drug Discovery and Development","volume":"13","author":"Chen","year":"2022","journal-title":"J. Cancer Ther."},{"key":"ref_64","doi-asserted-by":"crossref","unstructured":"Coelho, L.L., Vianna, M.M., da Silva, D.M., Gonzaga, B.M.d.S., Ferreira, R.R., Monteiro, A.C., Bonomo, A.C., Manso, P.P.d.A., de Carvalho, M.A., and Vargas, F.R. (2024). Spheroid Model of Mammary Tumor Cells: Epithelial\u2013Mesenchymal Transition and Doxorubicin Response. Biology, 13.","DOI":"10.3390\/biology13070463"},{"key":"ref_65","doi-asserted-by":"crossref","unstructured":"Lin, Y.N., Nasir, A., Camacho, S., Berry, D.L., Schmidt, M.O., Pearson, G.W., Riegel, A.T., and Wellstein, A. (2020). Monitoring cancer cell invasion and t-cell cytotoxicity in 3d culture. J. Vis. Exp., 160.","DOI":"10.3791\/61392-v"},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"4775","DOI":"10.1007\/s13346-025-01896-8","article-title":"Immunostimulatory effects of IL-12 targeted pH-responsive nanoparticles in macrophage-enriched 3D immuno-spheroids in vitro model","volume":"15","author":"Silveira","year":"2025","journal-title":"Drug Deliv. Transl. Res."},{"key":"ref_67","doi-asserted-by":"crossref","first-page":"454","DOI":"10.1177\/2211068216652846","article-title":"High-Throughput 3D Tumor Spheroid Screening Method for Cancer Drug Discovery Using Celigo Image Cytometry","volume":"22","author":"Kessel","year":"2017","journal-title":"SLAS Technol."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/8\/3529\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,4,25]],"date-time":"2026-04-25T04:15:31Z","timestamp":1777090531000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/27\/8\/3529"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2026,4,15]]},"references-count":67,"journal-issue":{"issue":"8","published-online":{"date-parts":[[2026,4]]}},"alternative-id":["ijms27083529"],"URL":"https:\/\/doi.org\/10.3390\/ijms27083529","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2026,4,15]]}}}