Related Experiment Video
Updated: Aug 18, 2026
Vitrification of Ovarian Cortex Tissue to Achieve a Glassy State of Aggregation
Published on: August 9, 2024
Coculture of monkey ovarian tissue increases survival after vitrification and slow-rate freezing
Richard R Yeoman1, Don P Wolf, David M Lee
1Division of Reproductive Sciences, Oregon National Primate Research Center, Oregon Health and Sciences University, 505 185th Avenue, Beaverton, OR 97006, USA. yeomanr@ohsu.edu
Objective:
To assess whether coculture of monkey ovarian tissue after low-temperature storage enhances follicular viability. To assess a novel method of vitrifying ovarian tissue.
Design:
Prospective in vitro study.
Setting:
University-affiliated national research center.
Animal(S):
Ovaries from 15 cynomolgus or rhesus macaques (1-11 years).
Intervention(S):
Vitrification using a containerless liquid nitrogen emersion system that involves dropping thin cortical pieces suspended in cyroprotectant directly into liquid nitrogen with outcome compared with slow-rate-controlled freezing. Before analysis, some of the thawed tissue was cocultured on mitotically inactivated mouse fetal fibroblast monolayers supplemented with FSH, insulin, transferrin, and selenium.
Main Outcome Measure(S):
Percentage of oocytes viable using live-dead fluorescent staining with carboxyfluorescein diacetate succinimidyl ester and propidium iodide.
Result(S):
Postthaw survival rates were 70.4% +/- 4.8% of 1,705 follicles after vitrification and 67.3% +/- 1.9% of 1,895 follicles after slow-rate freeze in six trials with each method. Coculture of the thawed tissue increased the viability, respectively, to 89% +/- 2.1% of 2,833 follicles previously vitrified and to 90.3% +/- 1.9% of 2,109 follicles after a slow-rate freeze (P<.01). Primordial follicles (30- to 50-microm diameter) were the vast majority of surviving follicles after thaw and coculture. Follicular viability in control fresh tissue (eight trials) was 76.0% +/- 4.1%, suggesting negligible loss in follicular viability after cryopreservation.
Conclusion(S):
Coculture of thawed ovarian tissue on mouse fetal fibroblasts and FSH increases the percentage of viable follicles. A novel method of vitrifying ovarian tissue is as effective as slow-rate freezing. These approaches may improve graft survival and function when used to treat chemotherapy-induced sterility.
