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Updated: Aug 16, 2025
Characterization of Human Monocyte-derived Dendritic Cells by Imaging Flow Cytometry: A Comparison between Two Monocyte Isolation Protocols
Published on: October 18, 2016
Guidelines for DC preparation and flow cytometric analysis of human lymphohematopoietic tissues
Lukas Heger1, Diana Dudziak1,2,3, Lukas Amon1
1Laboratory of Dendritic Cell Biology, Department of Dermatology, University Hospital Erlangen, Hartmannstraße 14, D-91052, Erlangen, Germany.
Insights
This guide offers detailed protocols for isolating and analyzing human dendritic cells (DC) from various 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 (DC) are crucial immune cells involved in initiating adaptive immune responses.
- Standardized protocols are needed for reliable DC preparation and analysis across different laboratories.
Purpose of the Study:
- To provide state-of-the-art protocols for the isolation, characterization, and functional analysis of human dendritic cells (DC).
- To facilitate the generation of single-cell suspensions and purification of primary human DC for downstream applications.
Main Methods:
- Detailed protocols for generating single-cell suspensions from human lymphohematopoietic tissues (blood, spleen, thymus, tonsils).
- Flow cytometry for phenotype analysis and cell sorting of primary human DC.
- Protocols adaptable for RNA sequencing, functional assays, and cellular enrichment.
Main Results:
- Established robust methods for preparing and analyzing human dendritic cells (DC) from diverse tissues.
- Enabled efficient flow cytometric analysis and cell sorting of primary human DC.
- Demonstrated the utility of prepared cells for various downstream molecular and functional studies.
Conclusions:
- This article serves as an essential resource for basic and clinical immunologists working with dendritic cells (DC).
- The provided protocols ensure high-quality data for DC 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 non-lymphoid tissues. Within this article, detailed protocols are presented that allow for the generation of single cell suspensions from human lymphohematopoietic tissues including blood, spleen, thymus, and tonsils with a focus on the subsequent analysis of DC via flow cytometry, as well as flow cytometric cell sorting of primary human DC. Further, prepared single cell suspensions as well as cell sorter-purified DC can be subjected to other applications including cellular enrichment procedures, RNA sequencing, functional assays, and many more. 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.
