Nasal turbinate epithelial cells were isolated from rats, rabbits, and humans using either a surgical or an in situ enzyme incubation technique. The culture conditions that permit optimal cell attachment and selective growth of the nasal epithelial cells were determined. These conditions will permit the long-term culture of these cells where typically 20 to 30 population doublings were observed. Differences between rat and human nasal epithelial cells were seen in substrate requirements, colony-forming efficiency, and response to fetal bovine serum and bovine serum albumin. These methodology and results will permit mechanistic studies of normal and abnormal cellular function and permit comparative response studies between nasal epithelial cells from rats and humans.