Guidelines for preparation and flow cytometry analysis of human nonlymphoid tissue DC
Diana Dudziak1,2, Lukas Heger2,3, William W Agace4,5
1Institute of Immunology, Jena University Hospital, Friedrich-Schiller-University, Jena, Germany.
Insights
This article provides detailed protocols for preparing and analyzing dendritic cells (DC) from human tissues using flow cytometry. These methods are essential for immunologists studying DC function in health and disease.
Area of Science:
- Immunology
- Cell Biology
Background:
- Dendritic cells (DCs) are crucial immune cells involved in initiating adaptive immune responses.
- Standardized protocols are needed for reliable DC isolation and analysis from diverse human tissues.
Purpose of the Study:
- To present state-of-the-art protocols for the preparation and flow cytometric analysis of human dendritic cells.
- To enable the generation of single-cell suspensions from various human nonlymphoid tissues and tumors.
Main Methods:
- Detailed protocols for generating single-cell suspensions from human lung, skin, gingiva, intestine, tumors, and tumor-draining lymph nodes.
- Flow cytometry techniques for phenotype analysis of dendritic cells.
Main Results:
- Established methods for preparing high-quality single-cell suspensions suitable for dendritic cell analysis.
- Protocols validated for use with various human tissues and lymphoid organs.
Conclusions:
- This article serves as an essential resource for basic and clinical immunologists.
- The provided protocols facilitate robust dendritic cell research and clinical applications.
Abstract:
This article is part of the Dendritic Cell Guidelines article series, which provides a collection of state-of-the-art protocols for the preparation, phenotype analysis by flow cytometry, generation, fluorescence microscopy, and functional characterization of mouse and human dendritic cells (DC) from lymphoid organs, and various nonlymphoid tissues. Within this article, detailed protocols are presented that allow for the generation of single-cell suspensions from human nonlymphoid tissues including lung, skin, gingiva, intestine as well as from tumors and tumor-draining lymph nodes with a subsequent analysis of dendritic cells by flow cytometry. Further, prepared single-cell suspensions can be subjected to other applications including cellular enrichment procedures, RNA sequencing, functional assays, etc. While all protocols were written by experienced scientists who routinely use them in their work, this article was also peer-reviewed by leading experts and approved by all co-authors, making it an essential resource for basic and clinical DC immunologists.
