Sphingosine-1-phosphate protects human ovarian follicles from apoptosis in vitro

We aimed to analyze if anti-apoptotic agent sphingosine-1-phosphate offers protection against in vitro follicle atresia during culture of human ovarian cortical samples. A translational research study of ex-vivo and in-vitro models of human ovarian tissue. Ovarian cortical tissue fragments (1 × 0.5 cm) were obtained from young patients (n = 15 mean age ± SD: 29.4 ± 2.5) undergoing laparoscopic excision of benign ovarian cysts. The samples were cultured for 4 days in 24-well format culture plate using conventional culture techniques. S1P was added to culture media at 200 and 400 μM concentrations. At the end of culture period the samples were processed for both histomorphological assessment and detection of apoptosis with immunohistochemistry and western blot methods using apoptosis marker cleaved caspase-3. In vitro estradiol (E2) and AMH productions of the samples were measured with ELISA. Follicle counts were expressed as the mean number of follicles per mm2. The mean numbers of primordial and secondary follicles were 3.2 ± 0.4 and 0.7 ± 0.2 respectively, in the fresh fixed uncultured samples. After four days of culture their numbers were significantly decreased to 0.8 ± 0.2 (p