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Updated: May 10, 2026
Vitrification of Ovarian Cortex Tissue to Achieve a Glassy State of Aggregation
Published on: August 9, 2024
Antiapoptotic agent sphingosine-1-phosphate protects vitrified murine ovarian grafts
Yung-Chieh Tsai1, Chii-Ruey Tzeng, Chia-Woei Wang
11Department of Obstetrics and Gynecology, Chi-Mei Medical Center, Tainan, Taiwan.
Abstract:
Significant follicle loss from frozen ovarian grafts is unavoidable. The authors evaluated the protective effects of the antiapoptotic agent sphingosine-1-phosphate (S1P) on vitrified ovarian grafts. Three-week-old sexually immature female FVB mice were divided into 4 groups, fresh, control without S1P, 0.5 mmol/L S1P, and 2 mmol/L S1P. The ovaries were pretreated with S1P for 1 hour and then cryopreserved by modified vitrification. The frozen-thawed ovaries were autotransplanted under the back muscles of mice for 10 days. Expression of apoptosis-related genes encoding caspase 3 and c-Myc was analyzed in the vitrified ovaries and 10 days after transplantation using real-time quantitative polymerase chain reaction. To quantify the ovarian reserve, anti-Müllerian hormone (AMH) levels and follicles were measured in the 10-day vitrified ovarian grafts. Caspase 3 and c-Myc messenger RNA did not differ significantly in the 4 groups after vitrification but was significantly upregulated in the control group after transplantation. The AMH levels and primordial follicle pool were significantly higher in the S1P-treated groups than in the control group but lower than that in the fresh group. The S1P protects vitrified ovarian grafts from ischemic reperfusion injury rather than from vitrification-associated process.
Insights
Sphingosine-1-phosphate (S1P) protects frozen ovarian grafts from damage after transplantation. This antiapoptotic agent helps preserve ovarian reserve by reducing injury from blood flow interruption, not the freezing process itself.
Area of Science:
- Reproductive Biology
- Cryobiology
- Oncology Supportive Care
Background:
- Ovarian tissue cryopreservation is crucial for fertility preservation, especially for cancer patients.
- Significant loss of ovarian follicles occurs during the process of cryopreservation and transplantation.
- Ischemic reperfusion injury after transplantation further damages ovarian grafts.
Purpose of the Study:
- To evaluate the protective effects of sphingosine-1-phosphate (S1P) on vitrified ovarian grafts.
- To determine if S1P mitigates damage associated with cryopreservation or post-transplantation ischemia.
Main Methods:
- Ovarian tissues from immature female mice were pretreated with varying concentrations of S1P (0.5 mmol/L, 2 mmol/L) or vehicle.
- Tissues underwent modified vitrification, followed by autotransplantation under the back muscles for 10 days.
- Apoptosis-related gene expression (caspase 3, c-Myc), anti-Müllerian hormone (AMH) levels, and follicle counts were analyzed.
Main Results:
- S1P treatment did not significantly alter apoptosis-related gene expression immediately after vitrification.
- Following transplantation, S1P-treated groups showed significantly lower upregulation of caspase 3 and c-Myc compared to controls.
- AMH levels and primordial follicle counts were significantly higher in S1P-treated grafts than controls, but lower than fresh, untransplanted ovaries.
Conclusions:
- Sphingosine-1-phosphate (S1P) demonstrates protective effects on vitrified ovarian grafts.
- S1P primarily protects against ischemic reperfusion injury occurring after transplantation, rather than the vitrification process itself.
- S1P represents a potential therapeutic agent to improve ovarian reserve and graft survival in fertility preservation.
