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.

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

Stphanie 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 Rene Van Amerongen

Part IV: TRANSLATIONAL APPLICATIONS

Laoighse Mulrane, William M. Galla