CLONAL GROWTH OF EPITHELIAL-CELLS FROM NORMAL ADULT HUMAN BRONCHUS

Abstract

Normal primary epithelial cell cultures devoid of fibroblastic cells were developed from tissue explants of adult human bronchi. Conditions for clonal growth of secondary cultures of bronchial epithelial cells were optimized by coculturing the human cells with mitomycin C growth-arrested Swiss 3T3 mouse feeder cells, lowering the Ca concentration of medium M199, and supplementing it with hydrocortisone, insulin, cholera toxin, epidermal growth factor and 1.25% fetal bovine serum. The epithelial cells grew for an average of 35 population doublings and had the normal human karyotype, expressed keratin and blood group antigen epithelial cell markers, metabolized benzo(a)pyrene and were capable of differentiating into ciliated and squamous cells. This culture system makes it potentially possible to investigate various aspects of differentiation and carcinogenesis in human bronchial epithelial cells.