Abstract:

BACKGROUND: Studies demonstrated that Sertoli cells have immunosuppression, which can provide immune privilege for transplanted cells in vitro. Previous researches indicated that Sertoli cells-secreted active compounds have promotive effects on other cells. However, there is not a complete method for Sertoli cells purification.
OBJECTIVE: To discuss a method for isolating Sertoli cells with high purity.
METHODS: The testicular seminiferous tubules were isolated from 20-day old male SD rats, digested by enzyme and were plated onto poly-L-lysine (PLL)-coated tissue culture dishes followed by 7 days culture, and then incubated with Tris-HCL. Purity of Sertoli cells were identified by immunohistochemistry of FasL with a counterstain of Hoechst33342.
RESULTS AND CONCLUSION: Most Sertoli cells began to stick on the flasks with round or oval shapes at 24 hours after culture, and many germ cells could be seen floating on the liquid. The cytoplasty become greater and refraction turned to weakening at 48 hours after culture. After 4-6 days, cells were fully spread and become monolayer. And the mass to charge ratio was (7-9):1, the shape and proportion of the cells had no change. The purity of the Sertoli cells was (95.64±2.76)%. The results demonstrated that this method is a relatively easy, cheap, stable, and efficient for isolating Sertoli cells.