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This second edition volume provides new and updated chapters detailing simple and accessible experiment protocols to explore thymus biology. Chapters are divided into three parts presenting short reviews, analysis strategies, protocols for cell preparation, flow cytometry analyses, Innate Lymphoid Cells (ILC), mouse T-cell development, antigen receptor-less cousins of T cells, bone marrow chimeras, thymic stroma, and multiple aspects of thymocyte biology. Written in the 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 protocols, and notes on troubleshooting and avoiding known pitfalls.
Authoritative and cutting-edge, T-Cell Development: Methods and Protocols, Second Edition aims to be a useful practical guide to help readers overcome obstacles associated with experimental approaches of T-cell development.
List of contents
A beginner's guide to T cell development.- Revelations in thymic epithelial cell biology and heterogeneity from single cell RNA sequencing and lineage tracing methodologies.- Early development of innate lymphoid cells.- Development of gd T cells, Soldiers on the Front Lines of Immune Battles.- Principles of Advanced Flow Cytometry: A Practical Guide.- Genetic strategies to study T cell development.- A simplified approach to evaluating T cell development by flow cytometry.- Purification of thymocyte and T cell subsets.- Generation of bone marrow chimeras.- Generation of murine T cell effector populations in vitro.- Large-scale isolation of mouse thymic epithelial cells.- Generation of retrogenic mice to investigate T cell developmentPurification of bone marrow precursors to T cells and ILCs.- Studying T cell Development in Neonatal and Adult Thymic Slices.- Induction of Human T Cell Development In Vitro with OP9-DL4-7FS Cells Expressing Human Cytokines.- Methods for Study of Mouse T Cell Receptor a and b Gene Rearrangements.- Ultrasound guided intra-thymic cell injection.- Fetal thymic organ culture and negative selection.- In Vitro Analysis of Thymocyte Signaling.- Identification and purification of human T cell precursors.- In vitro model systems to study human T cell development.- Zebrafish: A Tractable Model for Analysis of T cell Development.
