Isolation of murine thymic epithelium and an improved method for its propagation in vitro

A reliable and reproducible method for the isolation and propagation of thymic epithelial cells is described. Thymic epithelial cells from enzymatically dissociated thymus stroma are first enriched by separation on a discontinuous Percoll density gradient. The cell fractions enriched for epithelial cells are then cultured with irradiated fibroblasts in Ham's F-12 nutrient medium. Colonies of cells in these cultures contain keratin and exhibit morphologic characteristics of epithelial cells. When subcultured, the epithelial cells no longer require irradiated fibroblasts as filler cells. Some of the epithelial cells in vitro retain expression of class II (Ia) major histocompatibility antigens. The generation of defined cultures of thymic epithelial cells promises to be useful in defining their role in T cell differentiation.