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.
Highlights
Secondary lymphoid organs (SLO) are one of the hallmarks of the mammalian immune system
The stromal cell compartment provides a scaffold for these immune cell interactions, while forming a niche which is favorable for survival and, in case of an adaptive response, proliferation of hematopoietic cells
Cells were detached with trypsin and kept in a single cell suspension in PBS supplemented with 2% (v/v) heat inactivated (h.i.) new born calf serum (NBCS) on ice to prevent re-attaching
Summary
Secondary lymphoid organs (SLO) are one of the hallmarks of the mammalian immune system. The stromal cell compartment provides a scaffold for these immune cell interactions, while forming a niche which is favorable for survival and, in case of an adaptive response, proliferation of hematopoietic cells. This is done by offering structural support in combination with secretion of homeostatic and, when activated, pro-inflammatory mediators [2,3,4,5,6]. Advances made in the past decade to analyze the structured anatomy of SLO and to PLOS ONE | DOI:10.1371/journal.pone.0167555 December 1, 2016
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