This volume provides key molecular biological protocols and experimental strategies currently employed to study the various stages...
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This volume provides key molecular biological protocols and experimental strategies currently employed to study the various stages of mammary gland development. Chapters are divided into four parts covering genetically modified mouse models that exhibit prenatal and pubertal mammary gland phenotypes, 2D and 3D-model culture systems, methodologies for mammary stem cell isolation, and translational applications that provide a bridge between experimental studies of mammary gland development and the study of human breast cancer. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Mammary Gland Development: Methods and Protocols will be useful for researchers whose primary interest is in mammary gland development; developmental biologists, epithelial cell biologists, and those with an interest in molecular mechanisms underlying breast cancer.
Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts Includes supplementary material: sn.pub/extras Inhalt 1. Overview of the Development of the Mouse Mammary Gland; A Comparison of Mammary Gland Development in Mouse and Humans Torsten Stein and Sara McNally Part I: In Vivo Models 2. Prenatal Mammary Gland Development in the Mouse - Research Models and Techniques for its Study from Past to Present Jaqueline Veltmaat 3. Pubertal Mammary Gland Development: Elucidation Of In Vivo Morphogenesis Using Murine Models Jean Mcbryan and Jillian Howlin 4. Analysis of Mammary Gland Phenotypes by Transplantation of the Genetically Marked Mammary Epithelium Duje Buric and Cathrin Brisken 5. Pubertal Ductal Morphogenesis: Isolation and Transcriptome Analysis of the Terminal End Bud Joanna S. Morris and Torsten Stein 6. RNA Profiling of Non-Cultured Fibroblasts Isolated from Pubertal Mouse Mammary Gland Sections Ayman M. Ibrahim, Claire Cairney, Joanna S. Morris, and Torsten Stein 7. Analysis of the Involuting Mouse Mammary Gland: An In Vivo Model for Cell Death Bethan Lloyd-Lewis, Timothy J. Sargeant, Peter A. Kreuzaler, Henrike K. Resemann, Sara Pensa, and Christine J. Watson Part II: In Vitro 2D & 3DModels 8. Contractility Assay for Established Myoepithelial Cell Lines Stéphanie Cagnet, Marina A. Glukhova, and Karine Raymond 9. The use of 3D Culture of Primary Mammary Epithelial Cells to Define Molecular Entities Required for Acinus Formation. Analysing MAP Kinase Phosphatases Malgorzata Gajewska and Sara Mcnally 10. A 3D Fibroblast-Epithelium Co-culture Model for Understanding Micro-Environmental Role in Branching Morphogenesis of the Mammary Gland Zuzana Koledova and Pengfei Lu 11. RNA Sequencing Analysis Reveals Expression of a Transient EMT Profile During the Early During Organization of MCF10A Cells in 3D Benedikt Minke, Finian Martin, Walter Kolch, and Drieke Vandamme 12. A 3D Culture Model to Study How Fluid Pressure and Flow Affect the Behavior of Aggregates of Epithelial Cells Alexandra S. Piotrowski, Allison K. Simi, Mei Fong Pang, Joe Tien, and Celeste M. Nelson Part III: MAMMARY STEM CELLS 13. Purification of Distinct Subsets of Epithelial Cells from Normal Human Breast Tissue Mona Shehata and John Stingl 14. Techniques for the Reprogramming of Exogenous Stem/Progenitor Cell Populations Towards a Mammary Epithelial Cell Fate Gilbert Smith and Corrine A. Boulanger 15. Lineage Tracing of Mammary Stem and Progenitor Cells< Anoeska A.A. Van De Moosdijk, Nai Yang Fu, Anne C. Rios, Jane E. Visvader, and Renée Van Amerongen Part IV: TRANSLATIONAL APPLICATIONS 16. Assessment of Novel Proteins by Tissue Microarray Technology Laoighse Mulrane, William M. Gallagher, and Darran P. O'Connor 17. Patient-Derived Xenografts of Breast Cancer Damir Varelija, Christopher Byrne, Sinead Cocchiglia, and Leonie Young