The field of mammary gland biology and breast cancer research encompasses a wide range of topics and scientific questions, which span domains of molecular, cell and developmental biology, cancer research, and veterinary and human medicine, with interdisciplinary overlaps to non-biological domains. Accordingly, mammary gland and breast cancer researchers employ a wide range of molecular biology methods, in vitro techniques, in vivo approaches as well as in silico analyses. The list of techniques is ever-expanding; together with the refinement of established, staple techniques in the field, new technologies keep emerging thanks to technological advances and scientific creativity. This issue of the Journal of Mammary Gland Biology and Neoplasia represents a compilation of original articles and reviews focused on methods used in mammary gland biology and breast cancer research.

Department of Histology and Embryology Faculty of Medicine Masaryk University Brno Czech Republic

Department of Pathology University of California San Francisco USA

Stem Cell Research Unit Department of Anatomy Faculty of Medicine School of Health Sciences Biomedical Center University of Iceland Reykjavík Iceland

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Schultz-Pernice I, Engelbrecht LK, Petricca S, Scheel CH, Twigger A-J. Morphological Analysis of Human Milk Membrane Enclosed Structures Reveals Diverse Cells and Cell-like Milk Fat Globules. J Mammary Gland Biol Neoplasia. 2020 doi: 10.1007/s10911-020-09472-1. PubMed DOI PMC

Martin Carli JF, Trahan GD, Jones KL, et al. Single Cell RNA Sequencing of Human Milk-Derived Cells Reveals Sub-Populations of Mammary Epithelial Cells with Molecular Signatures of Progenitor and Mature States: a Novel, Non-invasive Framework for Investigating Human Lactation Physiology. J Mammary Gland Biol Neoplasia. 2020 doi: 10.1007/s10911-020-09466-z. PubMed DOI PMC

Twigger A-J, Engelbrecht LK, Bach K, Schultz-Pernice I, Petricca S, Scheel CH, et al. Transcriptional changes in the mammary gland during lactation revealed by single cell sequencing of cells from human milk. bioRxiv. 2020;2020.11.06.371443. PubMed PMC

Stewart TA, Davis FM. Got Milk? Identifying and Characterizing Lactation Defects in Genetically-Engineered Mouse Models. J Mammary Gland Biol Neoplasia. 2020 doi: 10.1007/s10911-020-09467-y. PubMed DOI

Feigman MJ, Moss MA, Chen C, Cyrill SL, Ciccone MF, Trousdell MC, et al. Pregnancy reprograms the epigenome of mammary epithelial cells and blocks the development of premalignant lesions. Nat Commun. 2020;11:2649. doi: 10.1038/s41467-020-16479-z. PubMed DOI PMC

Ciccone MF, Trousdell MC, dos Santos CO. Characterization of Organoid Cultures to Study the Effects of Pregnancy Hormones on the Epigenome and Transcriptional Output of Mammary Epithelial Cells. J Mammary Gland Biol Neoplasia. 2020 doi: 10.1007/s10911-020-09465-0. PubMed DOI PMC

Sumbal J, Chiche A, Charifou E, Koledova Z, Li H. Primary Mammary Organoid Model of Lactation and Involution. Front Cell Dev Biol [Internet]. Frontiers; 2020 [cited 2020 Apr 5];8. Available from: https://www.frontiersin.org/articles/10.3389/fcell.2020.00068/full. PubMed PMC

Sumbal J, Budkova Z, Traustadóttir GÁ, et al. Mammary Organoids and 3D Cell Cultures: Old Dogs with New Tricks. J Mammary Gland Biol Neoplasia. 2020 doi: 10.1007/s10911-020-09468-x. PubMed DOI

Wrenn ED, Moore BM, Greenwood E, et al. Optimal, Large-Scale Propagation of Mouse Mammary Tumor Organoids. J Mammary Gland Biol Neoplasia. 2020 doi: 10.1007/s10911-020-09464-1. PubMed DOI PMC

Lan Q, Mikkola ML. Protocol: Adeno-Associated Virus-Mediated Gene Transfer in Ex Vivo Cultured Embryonic Mammary Gland. J Mammary Gland Biol Neoplasia. 2020 doi: 10.1007/s10911-020-09461-4. PubMed DOI PMC

Bu W, Li Y. Intraductal Injection of Lentivirus Vectors for Stably Introducing Genes into Rat Mammary Epithelial Cells in Vivo. J Mammary Gland Biol Neoplasia. 2020 doi: 10.1007/s10911-020-09469-w. PubMed DOI PMC

Hillers-Ziemer LE, Arendt LM. Weighing the Risk: effects of Obesity on the Mammary Gland and Breast Cancer Risk. J Mammary Gland Biol Neoplasia. 2020;25:115–131. doi: 10.1007/s10911-020-09452-5. PubMed DOI PMC

Giles ED, Wellberg EA. Preclinical Models to Study Obesity and Breast Cancer in Females: Considerations, Caveats, and Tools. J Mammary Gland Biol Neoplasia. 2020 doi: 10.1007/s10911-020-09463-2. PubMed DOI PMC

Wortzel I, Dror S, Kenific CM, Lyden D. Exosome-Mediated Metastasis: Communication from a Distance. Dev Cell. 2019;49:347–360. doi: 10.1016/j.devcel.2019.04.011. PubMed DOI

Pelissier Vatter FA, Lucotti S, Zhang H. Recent Advances in Experimental Models of Breast Cancer Exosome Secretion, Characterization and Function. J Mammary Gland Biol Neoplasia. 2020 doi: 10.1007/s10911-020-09473-0. PubMed DOI

Turashvili G, Brogi E. Tumor Heterogeneity in Breast Cancer. Front Med (Lausanne) [Internet]. 2017 [cited 2020 Dec 7];4. Available from: https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5727049/ PubMed PMC

Baek M, Chang JT, Echeverria GV. Methodological Advancements for Investigating Intra-tumoral Heterogeneity in Breast Cancer at the Bench and Bedside. J Mammary Gland Biol Neoplasia. 2020 doi: 10.1007/s10911-020-09470-3. PubMed DOI PMC

Swann JB, Smyth MJ. Immune surveillance of tumors. J Clin Invest. 2007;117:1137–1146. doi: 10.1172/JCI31405. PubMed DOI PMC

Mori H, Bolen J, Schuetter L, Massion P, Hoyt CC, VandenBerg S, Esserman L, Borowsky AD, Campbell MJ. Characterizing the tumor immune microenvironment with tyramide based multiplex immunofluorescence. J Mammary Gland Biol Neoplasia. 2021. PubMed PMC

van de Grift YBC, Heijmans N, van Amerongen R. How to Use Online Tools to Generate New Hypotheses for Mammary Gland Biology Research: A Case Study for Wnt7b. J Mammary Gland Biol Neoplasia. 2020 doi: 10.1007/s10911-020-09474-z. PubMed DOI PMC

Evolution and Self-renewal of the Journal of Mammary Gland Biology and Neoplasia