title: A Reproducible Method for Isolation and In Vitro Culture of Functional Human Lymphoid Stromal Cells from Tonsils.
abstract: The stromal compartment of secondary lymphoid organs is classicaly known for providing a mechanical scaffold for the complex interactions between hematopoietic cells during immune activation as well as for providing a niche which is favorable for survival of lymphocytes. In recent years, it became increasingly clear that these cells also play an active role during such a response. Currently, knowledge of the interactions between human lymphoid stroma and hematopoietic cells is still lacking and most insight is based on murine systems. Although methods to isolate stromal cells from tonsils have been reported, data on stability in culture, characterization, and functional properties are lacking. Here, we describe a reproducible and easy method for isolation and in vitro culture of functional human lymphoid stromal cells from palatine tonsils. The cells isolated express markers and characteristics of T cell zone fibroblastic reticular cells (FRCs) and react to inflammatory stimuli by upregulating inflammatory cytokines and chemokines as well as adhesion molecules, as previously described for mouse lymphoid stroma. Also, cultured tonsil stromal cells support survival of human innate lymphoid cells, showing that these stromal cells can function as bone fide FRCs, providing a favorable microenvironment for hematopoietic cells.
authors.affiliation: Department of Molecular Cell Biology and Immunology, VU University Medical Center, Amsterdam, The Netherlands.
authors.forename: Yotam E
authors.initials: YE
authors.lastname: Bar-Ephraim
authors.affiliation: Department of Molecular Cell Biology and Immunology, VU University Medical Center, Amsterdam, The Netherlands.
authors.forename: Tanja
authors.initials: T
authors.lastname: Konijn
authors.affiliation: Department of Otolaryngology, MC Hospital, Amsterdam, The Netherlands.
authors.forename: Mehmet
authors.initials: M
authors.lastname: Gönültas
authors.affiliation: Department of Molecular Cell Biology and Immunology, VU University Medical Center, Amsterdam, The Netherlands.
authors.forename: Reina E
authors.initials: RE
authors.lastname: Mebius
authors.affiliation: Department of Molecular Cell Biology and Immunology, VU University Medical Center, Amsterdam, The Netherlands.
authors.forename: Rogier M
authors.identifier: http://orcid.org/0000-0001-7471-7715
authors.initials: RM
authors.lastname: Reijmers
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000235:genetics / Q000276:immunology; D015415:Biomarkers / Q000378:metabolism; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000276:immunology*; D002450:Cell Communication / Q000276:immunology*; D002469:Cell Separation / Q000379:methods*; D018920:Coculture Techniques; D016207:Cytokines / Q000235:genetics / Q000276:immunology*; D016326:Extracellular Matrix Proteins / Q000235:genetics / Q000276:immunology; D005347:Fibroblasts / Q000166:cytology / Q000276:immunology; D005434:Flow Cytometry; D015870:Gene Expression; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D051379:Mice; D014066:Palatine Tonsil / Q000166:cytology* / Q000276:immunology; D061251:Primary Cell Culture; D015398:Signal Transduction; D017154:Stromal Cells / Q000166:cytology* / Q000187:drug effects / Q000276:immunology; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D015703:Antigens, CD; D015415:Biomarkers; D015815:Cell Adhesion Molecules; D016207:Cytokines; D016326:Extracellular Matrix Proteins; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma