Among different mammalian tissues, testis is found to be one of the most sensitive organs to TCDD exposure. In this study, primary Sertoli cell culture was established. The purity of the cultured cells was verified using 3β-hydroxysteroid dehydrogenase, alkaline phosphatase as well as testosterone induction assays. Effects of TCDD in modulating the expression of CYP1A1, aromatase, secretory products (i.e. Müllerian inhibiting substance (MIS), 17β-estradiol (E 2) and lactate) and markers for cell-cell interaction (i.e. sertolin and testin) were then examined. Our data demonstrated that Sertoli cells exposed to 0.2-2000 pg/ml of TCDD showed a dose dependent induction of CYP1A1 mRNA. The minimal dose of activation was 2 pg/ml, which indicated that the cell was very sensitive to TCDD exposure. However, there was little or no detectable level CYP1A1 protein and EROD activities found. Dose-dependent inductions of aromatase transcript (200%) and E 2 (20%) secretion were measured. In addition there was a significant reduction (40%) of MIS mRNA. No detectable change in the level of secreted lactate was observed. Sertolin and testin, the gene makers for cell-cell interactions were differentially modulated upon TCDD treatment. Taken together, the results implicated that TCDD exposure might interfere with the normal Sertoli cell functions. Copyright © 2004 Elsevier Ireland Ltd. All rights reserved.