Guidelines for mouse and human DC generation.

Area of Science:

• Immunology
• Cell Biology

Background:

• Dendritic cells (DC) are crucial immune cells involved in initiating adaptive immune responses.
• Standardized and reliable protocols are essential for reproducible DC research and clinical applications.

Purpose of the Study:

• To provide comprehensive, state-of-the-art protocols for the generation and characterization of mouse and human dendritic cells.
• To offer practical guidance, including common pitfalls and expert tips, for researchers working with diverse DC populations.

Main Methods:

• Detailed protocols for preparing and generating mouse and human DCs from various sources like bone marrow (BM), HoxB8 cells, CD34+ cells, and peripheral blood monocytes.
• Methods encompass phenotype analysis via flow cytometry, fluorescence microscopy, and functional characterization.
• Specific protocols cover murine cDC1, cDC2, pDC generation using Flt3L, BM-derived DC with GM-CSF, and human DC subset generation from CD34+ cells and monocytes.

Main Results:

• Established protocols for generating specific murine and human DC subsets (cDC1, cDC2, pDC, MoDC).
• Included are methods for DC subset enrichment, CRISPR/Cas9 editing, and clinical-grade human DC production.
• Protocols are validated by experienced scientists and peer-reviewed by experts.

Conclusions:

• This article serves as an essential resource for basic and clinical immunologists requiring robust DC generation and analysis protocols.
• The provided guidelines facilitate reproducible research and advancement in dendritic cell immunology.