Main description:

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.

Contents:

Part I: Background Information

1 A beginner's guide to T cell development

Remy Bosselut

2 Revelations in thymic epithelial cell biology and heterogeneity from single cell RNA sequencing and lineage tracing methodologies

Abigail Morales-Sanchez, Susannah C Shissler, Jennifer E Cowan, and Avinash Bhandoola

3 Early development of innate lymphoid cells

Yi Ding, Christelle Harly, Arundhoti Das and Avinash Bhandoola

4 Development of gd T cells, Soldiers on the Front Lines of Immune Battles

Alejandra V. Contreras and David L. Wiest

5 Principles of Advanced Flow Cytometry: A Practical Guide

Shafiuddin Siddiqui and Ferenc Livak

Part II: Core Approaches and Strategies

6 Genetic strategies to study T cell development

Remy Bosselut

7 A simplified approach to evaluating T cell development by flow cytometry

Jan Y.M. Lee and Paul E. Love

8 Purification of thymocyte and T cell subsets

Andrea C. Carpenter, Laura B. Chopp, Jong Kyong Kim and Remy Bosselut

9 Generation of bone marrow chimeras

Yayi Gao and Remy Bosselut

10 Generation of murine T cell effector populations in vitro

Melanie S. Vacchio

Part III: Specific Techniques

11 Large-scale isolation of mouse thymic epithelial cells Izumi Ohigashi, Mami Matsuda-Lennikov, and Yousuke Takahama

12 Generation of retrogenic mice to investigate T cell development

Yayi Gao and Remy Bosselut

13 Purification of bone marrow precursors to T cells and ILCs.

Devin Kenney and Christelle Harly

14 Studying T cell Development in Neonatal and Adult Thymic Slices

Laura L. McIntyre, Lydia K. Lutes and Ellen A. Robey

15 Induction of Human T Cell Development In Vitro with OP9-DL4-7FS Cells Expressing Human Cytokines

Mahmood Mohtashami, Patrick Brauer, and Juan Carlos Zuniga-Pflucker

16 Methods for Study of Mouse T Cell Receptor a and b Gene Rearrangements

Danielle J. Dauphars, Glendon Wu, Craig H. Bassing,, and Michael S. Krangel

17 Ultrasound guided intra-thymic cell injection

Hristo Georgie, Laura B. Chopp and Kristin A. Hogquist

18 Fetal thymic organ culture and negative selection

Emma Teixeiro, Mark A. Daniels

19 In Vitro Analysis of Thymocyte Signaling

Emma Teixeiro, and Mark A. Daniels

20 Identification and purification of human T cell precursors

Inge Van de Walle, Nina Lambrechts, Anais Derveeuw, Marieke Lavaert, Juliette Roels and Tom Taghon

21 In vitro model systems to study human T cell development

Nina Lambrechts, Kai Ling Liang, Imke Velghe, Steven Strubbe, Anne-Catherine Dolens and Tom Taghon

22 Zebrafish: A Tractable Model for Analysis of T cell Development

Robert Sertori, Yong Zhang and David L. Wiest