Human Sertoli cells were isolated from the testes of NOA and OA patients by a two-step enzymatic digestion and differential plating. Sertoli cells were identified by RT-PCR and immumochemistry. Sertoli cells obtained from OA patients with normal spermatogenesis served as the control. Nodal signaling genes and proteins expression profiles were investigated by PCR, immunochemistry. CCK8 was adopted to demonstrate whether Nodal promotes the proliferation of human Sertoli cells. Quantitative-PCR was used to detect the genes expressions of Nodal signaling after treatment of Nodal and its receptors inhibitors SB431542.