Chapter 2 Identification and Culture of Human Bronchial Epithelial Cells

This chapter examines the identification and culture of human bronchial epithelial cells. The development of methods for culturing human bronchial epithelial cells would permit analysis of factors that influence their normal differentiation and of the effects of chemical carcinogens and other toxins on these important target cells in human lung disease. The proliferative and differentiative characteristics of human bronchial epithelial cells in primary culture are described. Although ciliary activity is observed in some epithelial cells in primary culture for as long as 6 months, the majority of these cells differentiated to a keratin-producing squamous epithelium. The labeling index of epithelial cells and fibroblasts in the outgrowth is determined in primary cultures that have been incubated for a period of 25 days. Outgrowths of bronchial cells are fixed and washed and are reacted by indirect immunofluorescence with rabbit antibodies against purified fibronectin from fresh human serum. The ciliary activity, detected by localized movement of culture medium over the epithelial cells in the explant and in the outgrowth, is observed in cultures maintained for at least 6 months.