Freeze-thawing intact human ovary with its vascular pedicle with a passive cooling device

Abstract

Objective: To test the feasibility of freezing intact human ovary using a passive cooling device.

Design: Prospective experimental study.

Setting: Academic research unit of the department of gynecology in a university hospital.

Patient(s): Ovaries were obtained from three women who were between 29 and 36 years of age.

Intervention(s): Ovarian perfusion with cryoprotective solution and slow freezing with a cryofreezing container (5100 Cryo 1 degrees C Freezing Container; Nalgene, VWR, Belgium). Rapid thawing of ovaries by perfusion and bathing with decreased sucrose gradient.

Main outcome measure(s): Viability of follicles, stromal cells, and vascular components was assessed with live-dead stains in freshly removed ovary, after cryoprotectant exposure before freezing, and after thawing. Histological morphology was assessed at these three different times.

Result(s): The percentage of live follicles was 99.4% in fresh tissue, 98.1% after cryoprotectant exposure, and 75.1% after thawing. Viability assessment showed live stromal cells and small vessels after thawing. On histological evaluation, the morphology of follicles and cortical and medullar tissue was similar in all three groups.

Conclusion(s): We described the cryopreservation, using an accessible protocol, of intact human ovary with its vascular pedicle and proved high survival rates of follicles, small vessels, and stromal cells and a normal histological structure in all the ovarian components after thawing using our protocol.