The collection on Methods and Models in Mammary Gland Biology and Breast Cancer Research showcases recent advances in tools and models that enhance our understanding of mammary gland development and breast cancer. This collection includes sixteen articles, collectively addressing approaches to investigate key aspects of mammary gland biology and tumorigenesis, including hormonal signaling, tissue architecture, tumor microenvironment, and species-specific mammary development. The issue highlights innovations such as optimized progesterone receptor reporters, improved menopause models, and 3D-printed mammary epithelial structures. It also features advancements in organoid-based studies, in situ labeling of epithelial proliferation in large animals, preclinical models for breast cancer prevention, and high-resolution imaging techniques. Methodologies for studying macrophage-cancer cell interactions and lysosomal function are provided as step-by-step protocols. Additionally, review articles provide insights into diverse mammalian organoid systems, rat mammary tumor models, and strategies for modeling breast cancer metastasis. Together, these contributions advance mammary gland research by refining experimental approaches, expanding model diversity, and fostering translational applications in breast cancer.

Institute of Biotechnology University of Helsinki Helsinki Finland

Institute of Molecular Genetics of the Czech Academy of Sciences Prague Czech Republic

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Aarts MT, Wagner M, van der Wal T, van Boxtel AL, van Amerongen R. A molecular toolbox to study progesterone receptor signaling. J Mammary Gland Biol Neoplasia. 2023;28:24. PubMed PMC

Brezak M, Koledova S, Z. Defective mammary epithelial outgrowth in Transgenic EKAREV-NLS mice: correction via Estrogen supplementation and genetic background modification. J Mammary Gland Biol Neoplasia. 2025;30:1. PubMed PMC

Brisken C, Scabia V. 90 YEARS OF PROGESTERONE: progesterone receptor signaling in the normal breast and its implications for cancer. J Mol Endocrinol. 2020;65:T81–94. PubMed

Chalmers SB, van der Wal T, Fre S, Jonkers J. Fourteenth annual ENBDC workshop: methods in mammary gland biology and breast Cancer. J Mammary Gland Biol Neoplasia. 2023;28:22. PubMed PMC

Chapman RS, Lourenco PC, Tonner E, Flint DJ, Selbert S, Takeda K, Akira S, Clarke AR, Watson CJ. Suppression of epithelial apoptosis and delayed mammary gland Involution in mice with a conditional knockout of Stat3. Genes Dev. 1999;13:2604–16. PubMed PMC

Hardwick LJA, Davies BP, Pensa S, Burge-Rogers M, Davies C, Baptista AF, Knott R, McCrone S, Po I, Strugnell E, B. W., et al. In the murine and bovine maternal mammary gland signal transducer and activator of transcription 3 is activated in clusters of epithelial cells around the day of birth. J Mammary Gland Biol Neoplasia. 2024;29:10. PubMed PMC

Koskinen LM, Nieminen L, Arjonen A, Guzmán C, Peurla M, Peuhu E. Spatial engineering of mammary epithelial cell cultures with 3D Bioprinting reveals growth control by branch point proximity. J Mammary Gland Biol Neoplasia. 2024;29:5. PubMed PMC

Lloyd-Lewis B, D’Angelo ME, Prowting NB, Wiseman BE, Sargeant TJ, Watson CJ. Methods for investigating STAT3 regulation of lysosomal function in mammary epithelial cells. J Mammary Gland Biol Neoplasia. 2024;29:11. PubMed PMC

Machiela MN, Hovey RC. Intramammary labeling of epithelial cell division. J Mammary Gland Biol Neoplasia. 2024;29:17. PubMed PMC

Mayer LP, Dyer CA, Eastgard RL, Hoyer PB, Banka CL. Atherosclerotic lesion development in a novel ovary-intact mouse model of perimenopause. Arterioscler Thromb Vasc Biol. 2005;25:1910–6. PubMed

Mazan A, Marusiak AA. Protocols for Co-Culture phenotypic assays with breast Cancer cells and THP-1-Derived macrophages. J Mammary Gland Biol Neoplasia. 2024;29:4. PubMed PMC

Molière S, Martinet A, Jaulin A, Lodi M, Chamaraux-Tran T-N, Alpy F, Bierry G, Tomasetto C. Fast ultrasound scanning is a rapid, sensitive, precise and Cost-Effective method to monitor tumor grafts in mice. J Mammary Gland Biol Neoplasia. 2024;29:2. PubMed PMC

Nicotra R, Lutz C, Messal HA, Jonkers J. Rat models of hormone Receptor-Positive breast Cancer. J Mammary Gland Biol Neoplasia. 2024;29:12. PubMed PMC

Nuckhir M, Withey D, Cabral S, Harrison H, Clarke RB. State of the Art modelling of the breast Cancer metastatic microenvironment: where are we?? J Mammary Gland Biol Neoplasia. 2024;29:14. PubMed PMC

Obr AE, Edwards DP. The biology of progesterone receptor in the normal mammary gland and in breast cancer. Mol Cell Endocrinol. 2012;357:4–17. PubMed PMC

Ortiz JR, Lewis SM, Ciccone M, Chatterjee D, Henry S, Siepel A, Santos D, C. O. Single-Cell transcription mapping of murine and human mammary organoids responses to female hormones. J Mammary Gland Biol Neoplasia. 2024;29:3. PubMed PMC

Rafaeva M, Kaczyńska B, Redondo-Pedraza J, Bentires-Alj M, Khaled WT, Fre S. 15th annual ENBDC meeting: how do cellular potency, microenvironment and natural rhythms influence mammary gland biology and breast cancer?? J Mammary Gland Biol Neoplasia. 2024;29:19. PubMed

Rauner G. Using organoids to tap mammary gland diversity for novel insight. J Mammary Gland Biol Neoplasia. 2024;29:7. PubMed PMC

Saeki K, Ha D, Chang G, et al. Perimenopausal and menopausal mammary glands in a 4-vinylcyclohexene diepoxide mouse model. J Mammary Gland Biol Neoplasia. 2024;29:15. 10.1007/s10911-024-09569-x. PubMed PMC

Sumbal J, Budkova Z, Traustadóttir GÁ, Koledova Z. Mammary organoids and 3D cell cultures: old dogs with new tricks. J Mammary Gland Biol Neoplasia. 2020;25. PubMed

Wilby AJ, Cabral S, Zoghi N, Howell SJ, Farnie G, Harrison H. A novel preclinical model of the normal human breast. J Mammary Gland Biol Neoplasia. 2024;29:9. PubMed PMC