Sphingosine‐1‐phosphate reduces atresia of primordial follicles occurring during slow‐freezing and thawing of human ovarian cortical strips

Abstract

We aimed in this study to explore if sphingosine-1-phosphate (S1P) reduces apoptosis of primordial follicles during cryopreservation of human ovarian cortical samples. Ovarian cortical tissue fragments obtained from young patients who underwent laparoscopic excision of benign ovarian cysts were used for the experiments. The samples were slow-frozen and thawed with and without S1P at 200 and 400 μM, cultured for 1 day, and then were fixed and processed for both histomorphological assessment and detection of apoptosis with immunohistochemistry using apoptosis marker cleaved caspase-3. Follicle counts were expressed as the mean number of follicles per mm . The mean number of primordial follicles and in vitro estradiol (E2) and anti-mullerian hormone (AMH) production of the slow-frozen and thawed samples were significantly reduced compared with fresh unfrozen samples. S1P treatment at 400 μM but not 200 μM concentration resulted in a significant increase in the number of surviving primordial follicles and in vitro E2 and AMH productions of the samples compared with their counterparts slow-frozen without S1P. We found that that there was a significant decrease in the number of primordial follicles with their oocytes stained positive for cleaved caspase-3 in the slow-frozen samples S1P 400 μM in comparison with the samples slow-frozen without S1P. These results suggest that S1P may ameliorate follicle atresia occurring in human ovarian cortical samples during cryopreservation.

Female Fertility Preservation through Stem Cell-based Ovarian Tissue Reconstitution In Vitro and Ovarian Regeneration In Vivo

“…49,50 In addition and of direct relevance to clinical studies focused on restoration of fertility in women through orthotopic or heterotopic transplantation of cryopreserved-and-thawed ovarian cortical tissue strips, [21][22][23][24][25][26][27] S1P has also been reported to minimize the loss of primordial follicles that occurs during vitrification and thawing of mouse ovarian tissue grafts 51 as well as during the slow-freezing and thawing of human ovarian cortical tissue. 52 These encouraging outcomes have prompted efforts to identify additional factors capable of minimizing ovarian damage caused by anticancer treatments, with one of the most recent being a preclinical study in rats reporting that injections of curcumin and capsaicin can offset cyclophosphamideinduced premature ovarian failure. 53 In other studies, coadministration of imatinib (Gleevec), a 2-phenyl amino pyrimidine derivative that inhibits activity of the tyrosine kinase domains of c-Abl, c-Kit and platelet-derived growth factor receptor, has been reported to attenuate follicle depletion in mice caused by cisplatin treatment, 54 the latter of which activates apoptosis in mouse oocytes through the TAp63 signaling pathway.…”

“…49,50 In addition and of direct relevance to clinical studies focused on restoration of fertility in women through orthotopic or heterotopic transplantation of cryopreserved-and-thawed ovarian cortical tissue strips, 21–27 S1P has also been reported to minimize the loss of primordial follicles that occurs during vitrification and thawing of mouse ovarian tissue grafts 51 as well as during the slow-freezing and thawing of human ovarian cortical tissue. 52…”

How to Support the In Vitro Culture of Human Ovarian Cells and Follicles? A Preliminary Step for Assembling an Artificial Ovary

“…It is proposed that phenol red free DMEMF12 medium is superior, because phenol red is a weak estrogen with obvious biological effects (19). Also, it has shown detrimental effect on cat ovarian cryopreserved tissue (16).…”

The Impact of Chemotherapy on the Ovaries: Molecular Aspects and the Prevention of Ovarian Damage

“…In a human ovarian xenograft model, S1P can block the human apoptotic follicle death induced by cyclophosphamide and doxorubicin and preserve the primordial follicle stockpile [34,35]. Moreover, S1P seems to reduce the atresia of primordial follicles that occurs during the slow freezing and thawing of human ovarian cortical strips, confirming its protective role [36]. Recently, ceramide 1 phosphate (C1P), another sphingolipid, was also found to be a potential ovarian protective agent as ovarian administration of this drug reduces the ovarian damage induced by cyclophosphamide and protects the ovarian reserve via the inhibition of apoptosis and improvement of stromal vasculature [37].…”