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.

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Inhalt

Protocol for studying embryonic mammary gland branching morphogenesis ex vivo.- Multidimensional fluorescence imaging of embryonic and postnatal mammary gland development.- Single-cell transcriptomic and epigenetic analyses of mouse mammary development starting with the embryo.- Computational methods to identify cell fate determinants, identity transcription factors and niche induced signaling pathways for stem cell research.- Differential proteomic analysis of complex mixtures by label free nLC MS/MS.- Orthotopic transplantation of mouse mammary epithelial cells.- Lineage tracing methods to study mammary epithelial hierarchies in vivo.- Lentiviral transduction of mammary epithelial cells.- Modification of Single Cells within Mouse Mammary Gland Derived Acini via Viral Transduction.- Surgical Procedure for Implantation of Human Tumor Tissue into the Epithelium-free Mammary Fat Pad of Immunocompromised Mice to Generate Patient-Derived Xenografts (PDX).- The Chicken Embryo Chorioallantoic Membrane (CAM); a Versatile Tool for the Study of Patient-Derived Xenografts.- Intraductal injections into the mouse mammary gland.- Modelling breast cancer in organoid and intraductal models.- Single organoid droplet-based staining method for high-end 3D imaging of mammary organoids.- Divide and Conquer: Isolating Cell Populations to Investigate how Breast Cancer Risk Factors Alter the Breast Microenvironment.- An organotypic assay to study epithelial-fibroblast interactions in human breast.- Patient-derived explant cultures of normal and tumour human breast tissue.- Negative-Selection Enrichment of Circulating Tumor Cells from Peripheral Blood Using the Microfluidic CTC-iChip.- Measuring Mechanical Properties of Breast Cancer Cells with Atomic Force Microscopy.- Breast cancer stem cells: a clinician´s view.