Main description:

This second edition provides an overview of recent developments and approaches used by researchers to investigate the properties and functions of mammary epithelial and stem cells, which will contribute to understand the heterogeneity of the mammary gland and of breast cancer. Chapters detail processes used to characterize stem cells, single cell RNA sequencing, computational methods, sophisticated imaging techniques, and a variety of model systems, among others. 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 Stem Cells: Methods and Protocols, Second Edition aims to make available protocols used to navigate the intricate behavior of mammary stem cells and to gain further knowledge to take us closer to the design of innovative strategies to prevent and treat breast cancer.

Contents:

1. Protocol for studying embryonic mammary gland branching morphogenesis ex vivo

Qiang Lan, Jyoti Satta, Satu-Marja Myllymaki, Ewelina Trela, Riitta Lindstroem, Beata Kaczynska, Johanna Englund, and Marja L. Mikkola

2. Multidimensional fluorescence imaging of embryonic and postnatal mammary gland development

Claudia Carabana and Bethan Lloyd-Lewis

3. Single-cell transcriptomic and epigenetic analyses of mouse mammary development starting with the embryo

Zhibo Ma, Nikki K. Lytle, Cynthia Ramos, Razia F. Naeem, Geoffrey M. Wahl

4. Computational methods to identify cell fate determinants, identity transcription factors and niche induced signaling pathways for stem cell research

Muhammad Ali, Mariana Messias Ribeiro, and Antonio del Sol

5. Differential proteomic analysis of complex mixtures by label free nLC MS/MS

Iraide Escobes, Mikel Azkargorta, Ibon Iloro and Felix Elortza

6. Orthotopic transplantation of mouse mammary epithelial cells

Marisa M. Faraldo, Marina A. Glukhova, and Marie-Ange Deugnier

7. Lineage tracing methods to study mammary epithelial hierarchies in vivo

Veronica Rodilla and Silvia Fre

8. Lentiviral transduction of mammary epithelial cells

Richard Iggo

9. Modification of Single Cells within Mouse Mammary Gland Derived Acini via Viral Transduction

Lucia Garcia del Valle, Marta G. Montero, and Martin Jechlinger

10. Surgical Procedure for Implantation of Human Tumor Tissue into the Epithelium-free Mammary Fat Pad of Immunocompromised Mice to Generate Patient-Derived Xenografts (PDX)

Lacey E. Dobrolecki and Michael T. Lewis

11. The Chicken Embryo Chorioallantoic Membrane (CAM); a Versatile Tool for the Study of Patient-Derived Xenografts

Hugo Villanueva and Andrew G. Sikora

12. Intraductal injections into the mouse mammary gland

Erik Oliemuller, Richard Newman, and Beatrice A. Howard

13. Modelling breast cancer in organoid and intraductal models

Richard Iggo

14. Single organoid droplet-based staining method for high-end 3D imaging of mammary organoids

Jakub Sumbal and Zuzana Koledova

15. Divide and Conquer: Isolating Cell Populations to Investigate how Breast Cancer Risk Factors Alter the Breast Microenvironment

Lisa M. Arendt

16. An organotypic assay to study epithelial-fibroblast interactions in human breast

Jakub Sumbal, Thorarinn Gudjonsson, Gunnhildur Asta Traustadottir, and Zuzana Koledova

17. Patient-derived explant cultures of normal and tumour human breast tissue

Isabel Gris-Cardenas, Miriam Rabano, and Maria dM. Vivanco

18. Negative-Selection Enrichment of Circulating Tumor Cells from Peripheral Blood Using the Microfluidic CTC-iChip

Risa Burr, Jon F. Edd, Brian Chirn, Avanish Mishra, Daniel Haber, Mehmet Toner, and Shyamala Maheswaran

19. Measuring Mechanical Properties of Breast Cancer Cells with Atomic Force Microscopy

Barbara Zbiral, Andreas Weber, and Jose L. Toca-Herrera

20. Breast cancer stem cells: a clinicians view

Jose Schneider and Francisco J. Vizoso