Culture of rabbit pulmonary Clara cells

Rabbit pulmonary Clara cells isolated by centrifugal elutriation have been cultured for several weeks. Clara cells generally adhered poorly to plastic but the cells did attach to coated substrates. A selected medium supported serial subculture of Clara cells for 4-5 passages (1:2 split). The medium consisted of a basal nutrient medium, alpha MEM, supplemented with insulin, transferrin, epidermal growth factor, D-glucose, biotin, alpha-tocopherol, pituitary extract, trace elements and 2% Sephadex G-10-filtered FBS. Freshly prepared Clara cells showed high capacity to activate 2-aminofluorene (AF) to mutagenic products. However, after 6 weeks of culture the mutagenic activation of AF was reduced by 92.5% indicating loss of cytochrome P-450.