Abstract
A three-dimensional culture system for primary human mononuclear cells was developed, which reproducibly resulted in the formation of multicellular heterospheroids. Immunohistological characterization demonstrated not only the three-dimensional tissue-like aggregation of primary monocytes, B cells and T cells, but also the presence of macrophages and proliferating cells, indicating that a differentiation of monocytes to macrophages and an activation of cells were induced. Because of the phenotypical resemblance to granulomas the influence of an in vitro infection with mycobacteria on spheroid formation and morphology was analyzed. In comparison to control incubations, the formation of multinucleated giant cells and necrotic areas containing large numbers of mycobacteria could be observed, which resembled histological hallmarks of in situ tuberculoid granulomas. These characteristics have not been described for in vitro models of granuloma formation before and thus this new culture technique may prove to be a useful tool for analyzing aspects relevant to immunopathological processes in chronically inflamed tissues.
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