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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.
List of contents
Origin, Phenotype, and Function of Mouse Dendritic Cell Subsets.- Phenotypes and Functions of Human Dendritic Cell Subsets Particularly in the Tumor Microenvironment.- In Vivo Tracking of Dendritic Cell Migration.- In Vivo Analysis of Dendritic Cell Clonality.- In Vitro Generation of Murine Bone Marrow-Derived Dendritic Cells.- In Vitro Generation of Murine Dendritic Cells from Hoxb8-Immortalized Hematopoietic Progenitors.- In Vitro Generation of Murine CD8 + DEC205+ XCR1+ Cross-Presenting Dendritic Cells from Bone Marrow-Derived Hematopoietic Progenitors.- In Vitro Generation of Human Dendritic Cell Subsets from CD34+ Cord Blood Progenitors.- In Vitro Generation of Human Cross-Presenting Type 1 Conventional Dendritic Cells (cDC1s) and Plasmacytoid Dendritic Cells (pDCs).- Culture System Allowing the Simultaneous Differentiation of Human Monocytes into Dendritic Cells and Macrophages Using M-CSF, IL-4, and TNF-a.- Clonal Analysis of Human Dendritic Cell Progenitors Using a Stromal Cell Culture.- Enrichment of Large Numbers of Splenic Mouse Dendritic Cells after Injection of Flt3L-Producing Tumor Cells.- Isolation and Identification of Dendritic Cell Subsets from Human and Mouse Tumors.- Optimized Non-Viral Gene Disruption in Primary Murine and Human Myeloid Cells.- Monitoring the Interaction between Dendritic Cells and T Cells In Vivo with LIPSTIC.- Characterization of Dendritic Cell Metabolism by Flow Cytometry.- In Vivo and In Vitro Assay to Address Dendritic Cells Antigen Cross-Presenting Capacity.- Assays to Detect Cross-Dressing by Dendritic Cells In Vivo and In Vitro.- Assessing the Ability of Human Dendritic Cells to Stimulate Naive CD4+ and CD8+ T Cells.- In Vitro and In Vivo Assays to Evaluate Dendritic Cell Phagocytic Capacity.- Tracking Plasmacytoid Dendritic Cell Response to Physical Contact with Infected Cells.- Harnessing Single Cell RNA Sequencing to Identify Dendritic Cell Types, Characterize Their Biological States, and Infer Their Activation Trajectory.- Characterization of Developmental Trajectories of Dendritic Cell Hematopoiesis through Single Cell RNA Sequencing Methods.
