Human lymph node fibroblastic reticular cells maintain heterogeneous characteristics in culture
Janna E.G. Roet,
Andrew I. Morrison,
Aleksandra M. Mikula,
Michael de Kok,
Daphne Panocha,
Henk P. Roest,
Luc J.W. van der Laan,
Charlotte M. de Winde,
Reina E. Mebius
Affiliations
Janna E.G. Roet
Amsterdam UMC location Vrije Universiteit Amsterdam, Molecular Cell Biology & Immunology, De Boelelaan 1117, Amsterdam, the Netherlands; Amsterdam Institute for Immunology and Infectious diseases, Amsterdam, the Netherlands
Andrew I. Morrison
Amsterdam UMC location Vrije Universiteit Amsterdam, Molecular Cell Biology & Immunology, De Boelelaan 1117, Amsterdam, the Netherlands; Amsterdam Institute for Immunology and Infectious diseases, Amsterdam, the Netherlands
Aleksandra M. Mikula
Amsterdam UMC location Vrije Universiteit Amsterdam, Molecular Cell Biology & Immunology, De Boelelaan 1117, Amsterdam, the Netherlands; Amsterdam Institute for Immunology and Infectious diseases, Amsterdam, the Netherlands; Cancer Center Amsterdam, Cancer Biology & Immunology, Amsterdam, the Netherlands
Michael de Kok
Amsterdam UMC location Vrije Universiteit Amsterdam, Molecular Cell Biology & Immunology, De Boelelaan 1117, Amsterdam, the Netherlands; Amsterdam Institute for Immunology and Infectious diseases, Amsterdam, the Netherlands
Daphne Panocha
Amsterdam UMC location Vrije Universiteit Amsterdam, Molecular Cell Biology & Immunology, De Boelelaan 1117, Amsterdam, the Netherlands; Amsterdam Institute for Immunology and Infectious diseases, Amsterdam, the Netherlands; Cancer Center Amsterdam, Cancer Biology & Immunology, Amsterdam, the Netherlands
Henk P. Roest
Erasmus MC Transplant Institute, University Medical Center Rotterdam, Department of Surgery, Dr. Molewaterplein 40, 3015 GD Rotterdam, the Netherlands
Luc J.W. van der Laan
Erasmus MC Transplant Institute, University Medical Center Rotterdam, Department of Surgery, Dr. Molewaterplein 40, 3015 GD Rotterdam, the Netherlands
Charlotte M. de Winde
Amsterdam UMC location Vrije Universiteit Amsterdam, Molecular Cell Biology & Immunology, De Boelelaan 1117, Amsterdam, the Netherlands; Amsterdam Institute for Immunology and Infectious diseases, Amsterdam, the Netherlands; Cancer Center Amsterdam, Cancer Biology & Immunology, Amsterdam, the Netherlands; Corresponding author
Reina E. Mebius
Amsterdam UMC location Vrije Universiteit Amsterdam, Molecular Cell Biology & Immunology, De Boelelaan 1117, Amsterdam, the Netherlands; Amsterdam Institute for Immunology and Infectious diseases, Amsterdam, the Netherlands; Cancer Center Amsterdam, Cancer Biology & Immunology, Amsterdam, the Netherlands; Corresponding author
Summary: Fibroblastic reticular cells (FRCs) are mesenchymal stromal cells in human lymph nodes (LNs) playing a pivotal role in adaptive immunity. Several FRC subsets have been identified, yet it remains to be elucidated if their heterogeneity is maintained upon culture. Here, we established a protocol to preserve and culture FRCs from human LNs and characterized their phenotypic profile in fresh LN suspensions and upon culture using multispectral flow cytometry. We found nine FRC subsets in fresh human LNs, independent of donor, of which four persisted in culture throughout several passages. Interestingly, the historically FRC-defining marker podoplanin (PDPN) was not present on all FRC subsets. Therefore, we propose that CD45negCD31neg human FRCs are not restricted by PDPN expression, as we found CD90, BST1, and CD146/MCAM to be more widely expressed. Together, our data provide insight into FRC heterogeneity in human LNs, enabling further investigation into the function of individual FRC subsets.