VOLUME 2, ISSUE 3, P248-258, AUGUST 01, 2021
Yuting Fan, M.D., Ph.D., Colleen L. Flanagan, M.S.E., Margaret A. Brunette, M.S., Andrea S. Jones, M.S., Brendon M. Baker, Ph.D., Sherman J. Silber, M.D., Ariella Shikanov, Ph.D.
To evaluate the number and quality of ovarian follicles isolated from ovaries obtained from five deceased donors aged 18–26 years and to compare the follicle viability before and after cryopreservation.
Academic biomedical research laboratory.
Deidentified deceased human donors.
Slow-freeze cryopreservation and thawing.
Main Outcome Measure(s)
Follicle count, follicle density, follicle viability using immunohistochemical staining (terminal deoxynucleotidyl transferase dUTP nick end labeling).
The follicle density was negatively correlated with age in both cryopreserved/thawed and fresh groups. A total of 2,803 follicles from fresh and 1,608 follicles from cryopreserved tissues were classified and analyzed using the hematoxylin and eosin staining. There was no statistically significant difference in the percentage of morphologically normal follicles between the two groups. Terminal deoxynucleotidyl transferase dUTP nick end labeling assay indicated no higher DNA damage in the follicles and stroma cells after cryopreservation. Morphologically normal preantral follicles were enzymatically isolated from both fresh and cryopreserved tissues with 88.51 ± 5.93% (mean ± standard deviation) of the isolated follicles confirmed viable using LIVE/DEAD evaluation.
Our results indicate that the ovarian tissues from deceased donors maintain high quality after long-time extracorporeal circulation and transportation from the hospital to the laboratory. High survival rate of follicles at different developmental stages suggested tolerance to the cryopreservation process. Human ovarian tissues obtained from deceased donors are an ample source tissue and can be applied to promoting research and future clinical applications.