Main description:

This volume provides simple and accessible experiment protocols to explore thymus biology. T-Cell Development: Methods and Protocols is divided into three parts presenting short reviews on T cell development, analysis strategies, protocols for cell preparation, flow cytometry analyses, and multiple aspects of thymocyte biology. As a volume in the highly successful Methods in Molecular Biology series, chapters contain introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible protocols, and tips on troubleshooting and avoiding known pitfalls. Concise and easy-to-use, T-Cell Development: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Contents:

Part I: Background Information

1. 200 Million Thymocytes and I: A Beginner's Survival Guide to T cell Development

Melanie S. Vacchio, Thomas Ciucci and Remy Bosselut

2. Development of gd T Cells, the Special Forces Soldiers of the Immune System

David L. Wiest

Part II: Core Approaches and Strategies

3. Genetic Tools to Study T Cell Development

Thomas Ciucci, Melanie S. Vacchio and Remy Bosselut

4. Assessment of T cell Development by Flow Cytometry

Jan Y.M. Lee and Paul E. Love

5. Flow Cytometry Analysis of Thymic Cells and their Subpopulations

Izumi Ohigashi and Yousuke Takahama Epithelial

6. Identifying the Spatial Relationships of Thymic Stromal and Thymocyte Subsets by Immunofluorescence Analysis

Virginia Bain and Ellen R. Richie

7. Purification of Thymocyte and T Cell Subsets

Andrea C. Carpenter, Jong Kyong Kim, and Remy Bosselut

8. Retroviral Transduction of T Cells and T Cell Precursors

Amie Simmons and Jose Alberola-Ila

9. Bone Marrow and Fetal Liver Radiation Chimeras

Francis A Flomerfelt and Ronald E Gress

10. In vitro Analyses of T Cell Effector Differentiation

Elizabeth A. Wohlfert,

Andrea C. Carpenter, Yasmine Belkaid, and Remy Bosselut

Part III: Specific Techniques

11. Studying T Cell Development in Thymic Slices

Jenny O. Ross, Heather J. Melichar, Joanna Halkias, and Ellen A. Robey

12. FTOC-based Analysis of Negative Selection

Cody, A. Cunningham, Emma Teixeiro, and Mark A. Daniels

13. Reconstituted Thymus Organ Culture

Zimu Deng, Haifeng Liu, Jinxiu Rui, and Xiaolong Liu

14. Induction of T-cell Development in vitro by Delta-like (Dll)-Expressing Stromal Cells

Mahmood Mohtashami, Payam Zarin and Juan Carlos Zuniga-Pflucker

15. In vitro Analysis of Thymocyte Signaling

Cody A. Cunningham, Emma Teixeiro, Mark A. Daniels

16. Molecular Analysis of Mouse T Cell Receptor a and b Gene Rearrangements

Levi J. Rupp, Liang Chen, Michael S. Krangel, and Craig H. Bassing

17. Intrathymic Injection

Sugata Manna and Avinash Bhandoola

18. Analysis of Cell Proliferation and |Homeostasis using EdU Labeling

Francis A Flomerfelt and Ronald E Gress

19.

Characterization and Isolation of Human T Cell Progenitors

Karin Davids, Inge Van de Walle and Tom Taghon

20. Approaches to Study Human T Cell Development

Anne-Catherine Dolens, Inge Van de Walle and Tom Taghon

21. Humanized Mice to Study Human T Cell Development

Sarah Bonte, Sylvia Snauwaert, Stijn Vanhee, Anne-Catherine Dolens, Tom Taghon, Bart Vandekerckhove, and Tessa Kerre

22. Using the Zebrafish Model to Study T cell Development

Yong Zhang and David L. Wiest