Ciliogenesis and mucus synthesis in cultured human respiratory epithelial cells

The mechanisms for the regulation of ciliogenesis and for the synthesis of mucus are not well understood. We sought to develop a culture system for differentiating ciliated and secretory types of human respiratory epithelial (HRE) cells. Dissociated HRE cells obtained from nasal polyps and maxillary sinus mucosa were cultured on type I collagen gel. Cells grown to confluence on collagen gel lost their cilia and exhibited a flat, squamous-like appearance. After reaching confluence, the cultured cells with a collagen gel substrate were removed from plastic dishes and floated in the culture medium. After 7 days in the floating culture, some cells exhibited several centrioles or basal bodies, while others showed secretory granules. The secretory phenotype predominated after 7 days. After 14 days in the floating culture, nearly all cells were ciliated. The results demonstrate that the differentiation of HRE cells can be induced by floating cultured cells with a collagen gel substrate in a defined culture medium.