Main description:

This detailed volume provides methods that can be used to study dendritic cell (DC) ontogeny, isolation, migration, and functions. After an introduction to murine and human DC subsets and their unique transcriptional, phenotypic, and functional properties, the book continues with sections covering in vivo studies, in vitro differentiation, enrichment, functional characterization, as well as Omics approaches to study dendritic cells. Written for the highly successful Methods in Molecular Biology series, 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 practical, Dendritic Cells: Methods and Protocols is an ideal guide to familiarize readers with the current state of the art techniques to investigate these vital cells.

Contents:

Part I: Overview on Mouse and Human Dendritic Cells

1. Origin, Phenotype, and Function of Mouse Dendritic Cell Subsets

Dorothee Duluc and Vanja Sisirak

2. Phenotypes and Functions of Human Dendritic Cell Subsets Particularly in the Tumor Microenvironment

Candice Sakref, Nathalie Bendriss-Vermare, and Jenny Valladeau-Guilemond

Part II: In Vivo Study of Dendritic Cells

3. In Vivo Tracking of Dendritic Cell Migration

Michio Tomura

4. In Vivo Analysis of Dendritic Cell Clonality

Mar Cabeza-Cabrerizo

Part III: In Vitro Differentiation of Dendritic Cells

5. In Vitro Generation of Murine Bone Marrow-Derived Dendritic Cells

Yohan Gerber-Ferder, Pierre Bourdely, Mathias Vetillard, Pierre Guermonprez, and Julie Helft

6. In Vitro Generation of Murine Dendritic Cells from Hoxb8-Immortalized Hematopoietic Progenitors

Hans Hacker

7. In Vitro Generation of Murine CD8 + DEC205+ XCR1+ Cross-Presenting Dendritic Cells from Bone Marrow-Derived Hematopoietic Progenitors

Margaret E. Kirkling and Boris Reizis

8. In Vitro Generation of Human Dendritic Cell Subsets from CD34+ Cord Blood Progenitors

Pierre Bourdely, Roberto Savoldelli, Mathias Vetillard, Giorgio Anselmi, Julie Helft, and Pierre Guermonprez

9. In Vitro Generation of Human Cross-Presenting Type 1 Conventional Dendritic Cells (cDC1s) and Plasmacytoid Dendritic Cells (pDCs)

Xinlong Luo, Sreekumar Balan, Catharina Arnold-Schrauf, and Marc Dalod

10. Culture System Allowing the Simultaneous Differentiation of Human Monocytes into Dendritic Cells and Macrophages Using M-CSF, IL-4, and TNF-a

Javiera Villar, Alice Coillard, Corne van Roessel, and Elodie Segura

11. Clonal Analysis of Human Dendritic Cell Progenitors Using a Stromal Cell Culture

Kang Liu, Jaeyop Lee, and Thomas Luh

Part IV: Enrichment of Dendritic Cells

12. Enrichment of Large Numbers of Splenic Mouse Dendritic Cells after Injection of Flt3L-Producing Tumor Cells

Pauline Santa, Anais Roubertie, Severine Loizon, Anne Garreau, Amandine Ferriere, Dorothee Duluc, and Vanja Sisirak

13. Isolation and Identification of Dendritic Cell Subsets from Human and Mouse Tumors

Yamila Rocca, Aurelien Voissiere, Jenny Valladeau-Guilemond, and Nathalie Bendriss-Vermare

Part V: Functional Characterization of Dendritic Cells

14. Optimized Non-Viral Gene Disruption in Primary Murine and Human Myeloid Cells

Emily C. Freund, Simone M. Haag, Benjamin Haley, and Aditya Murthy

15. Monitoring the Interaction between Dendritic Cells and T Cells In Vivo with LIPSTIC

Giulia Pasqual, Aleksey Chudnovskiy, and Gabriel D. Victora

16. Characterization of Dendritic Cell Metabolism by Flow Cytometry

Eline C. Brombacher, Thiago A. Patente, Marjolein Quik, and Bart Everts

17. In Vivo and In Vitro Assay to Address Dendritic Cells Antigen Cross-Presenting Capacity

Pengju Ou, Lifen Wen, Hai Ni, and Cliff. Y. Yang

18. Assays to Detect Cross-Dressing by Dendritic Cells In Vivo and In Vitro

Alok Das Mohapatra and Pramod K. Srivastava

19. Assessing the Ability of Human Dendritic Cells to Stimulate Naive CD4+ and CD8+ T Cells

Alice Coillard, Tsing-Lee Tang-Huau, and Elodie Segura

20. In Vitro and In Vivo Assays to Evaluate Dendritic Cell Phagocytic Capacity

Lucie Maisonneuve and Benedicte Manoury

21. Tracking Plasmacytoid Dendritic Cell Response to Physical Contact with Infected Cells

Margarida Sa Ribeiro, Garima Joshi, Elodie Decembre, Celia Nuovo, Adrien Bosseboeuf, Alicia Bellomo, Manon Venet, Sonia Assil, and Marlene Dreux

Part VI: Omic Approaches to Study Dendritic Cells

22. Harnessing Single Cell RNA Sequencing to Identify Dendritic Cell Types, Characterize Their Biological States, and Infer Their Activation Trajectory

Ammar Sabir Cheema, Kaibo Duan, Marc Dalod, and Thien-Phong Vu Manh

23. Characterization of Developmental Trajectories of Dendritic Cell Hematopoiesis through Single Cell RNA Sequencing Methods

Regine J. Dress and Florent Ginhoux