Setup and Analysis of Human Tonsil and Adenoid-Based Immune Organoids for Studying T and B Cell-Mediated Adaptive Immunity.
- 1Medical Clinic III for Oncology, Hematology, Immuno-Oncology and Rheumatology, University Hospital Bonn, University of Bonn, Bonn, Germany.
- 2Department of Otorhinolaryngology, Head and Neck Surgery, University Hospital Bonn, University of Bonn, Bonn, Germany.
- 3Department of Otorhinolaryngology/Head and Neck Surgery, Johanniter-Waldkrankenhaus, Bonn, Germany.
Abstract
In recent years, demand for human-based in vitro models has grown substantially. In this regard, human secondary lymphoid organ (SLO)-derived ex vivo immune organoids are gaining recognition, as they provide a platform for studying highly differentiated, tissue-specific immune cell subsets found in SLOs, such as lymph nodes, tonsils, or spleen. These systems support the study of adaptive immune responses mediated by T and B cells, e.g., due to the presence of T follicular helper cells and germinal center B cells within these immune organoids. Here, we describe procedures for the isolation of human tonsil- or adenoid-derived immune cells, their cryopreservation and thawing, the establishment of immune organoids in vitro, and a detailed flow cytometry protocol for longitudinal phenotypic profiling of the immune organoids. In summary, these protocols serve as a foundation for downstream functional assays and complementary analyses in a human-first approach. © 2026 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Cell isolation from human tonsils and adenoids Basic Protocol 2: Thawing of cryopreserved tonsil and adenoid cells and establishment of immune organoids Basic Protocol 3: Flow cytometry for longitudinal phenotypical profiling of human tonsil/adenoid-derived immune organoids.
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BibTeX
@article{moll2026setup,
title = {{Setup and Analysis of Human Tonsil and Adenoid-Based Immune Organoids for Studying T and B Cell-Mediated Adaptive Immunity.}},
author = {Maximilian Moll and Klaus W Eichhorn and Thorsten Send and Dirk Baumjohann},
journal = {Current protocols},
year = {2026},
month = oct,
doi = {10.1002/cpz1.70485},
url = {https://europepmc.org/article/MED/42925496},
note = {CC BY},
}
RIS
TY - JOUR
TI - Setup and Analysis of Human Tonsil and Adenoid-Based Immune Organoids for Studying T and B Cell-Mediated Adaptive Immunity.
AU - Maximilian Moll
AU - Klaus W Eichhorn
AU - Thorsten Send
AU - Dirk Baumjohann
PY - 2026
DA - 2026/10/01
JO - Current protocols
DO - 10.1002/cpz1.70485
UR - https://europepmc.org/article/MED/42925496
AB - In recent years, demand for human-based in vitro models has grown substantially. In this regard, human secondary lymphoid organ (SLO)-derived ex vivo immune organoids are gaining recognition, as they provide a platform for studying highly differentiated, tissue-specific immune cell subsets found in SLOs, such as lymph nodes, tonsils, or spleen. These systems support the study of adaptive immune responses mediated by T and B cells, e.g., due to the presence of T follicular helper cells and germinal center B cells within these immune organoids. Here, we describe procedures for the isolation of human tonsil- or adenoid-derived immune cells, their cryopreservation and thawing, the establishment of immune organoids in vitro, and a detailed flow cytometry protocol for longitudinal phenotypic profiling of the immune organoids. In summary, these protocols serve as a foundation for downstream functional assays and complementary analyses in a human-first approach. © 2026 The Author(s). Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Cell isolation from human tonsils and adenoids Basic Protocol 2: Thawing of cryopreserved tonsil and adenoid cells and establishment of immune organoids Basic Protocol 3: Flow cytometry for longitudinal phenotypical profiling of human tonsil/adenoid-derived immune organoids.
SN - 2691-1299
ER -