Objective: To simplify the procedures of isolation of rat Sertoli cells and increase the yield of Sertoli cells cultured.
Methods: Decapsulated rat testis from 16-22-day-old SD rats were digested by sequential two-step digestion using 0.25% pancreatin, 0.05% collagenase in order, and then the cell suspension was cultured in incubator under the condition of 37 degrees C and 5% CO2 for 48 h. The yields were observed and Fas Ligand (Fas-L) was tested by immunohistochemical method.
Results: By the use of this modified method, the yielded sertoli cells accounted for 90% of all the cells harvested. High level expression of Fas-L was detected.
Conclusion: This modified method increases the yield of Sertoli cells and the procedures of isolation and culture are greatly simplified.