Infection of polarized human endometrial-gland epithelial cells obtained at hysterectomy with Chlamydia trachomatis serovar E may provide a relevant in vitro model for studies of pharmacokinetics in genital chlamydial infections. The minimal bactericidal concentration of azithromycin against C. trachomatis was lower in this model than in studies with nonpolarized cells (0.125 and 0.5 mg/L, respectively). Polarized cells also internalized more azithromycin over 24 hours. The results indicate that the eradication of chlamydial infections may be difficult to prove by antigen detection methods: the persistence of chlamydial envelope material within the intracellular vacuoles of azithromycin-treated cells may lead to a false-positive diagnosis of persisting chlamydial infection.