title: Viability of human ovarian tissue confirmed 5 years after freezing with spontaneous ice-formation by autografting and chorio-allantoic membrane culture.
abstract: To achieve optimal and uniform outcomes, slow cooling protocols for human ovarian tissues generally initiate ice formation at high sub-zero temperatures (-6 to -9 °C). The aim of the study was to investigate the function of ovarian tissue that had unintentionally self seeded at -20 °C during the freezing step, by examining its development following chicken embryonic chorioallantoic membrane (CAM) grafting and after transplantation back to the patient. Ovarian tissue was frozen in 6% (v/v) dimethyl sulfoxide, 6% (v/v) ethylene glycol and 0.15M sucrose which had self-seeded at -20 °C. Five years after cryopreservation, 8 pieces were thawed and transplanted back to the patient. Two small (1 × 2 × 1 mm) pieces of this thawed tissue were cultured in a CAM-system for 5 days to assess the tissue viability. The autografted ovarian tissue re-established spontaneous menstrual bleeding within five months and raised serum 17-β Estradiol from 19 to 330 pg/ml. Ultrasound revealed a dominant follicle at the site of the transplanted tissue in the follicular phase after the menstrual bleed. Analysis of the CAM cultured tissue established that 88% of the primordial follicles are degenerated and there was limited in growth of blood vessels. In conclusion, in spite of the damage caused by the cryopreservation with spontaneous ice-formation the viability could be confirmed by CAM culture and the restoration of ovarian function after auto-transplantation.
authors: Department of Obstetrics and Gynaecology, University Maternal Hospital, Cologne University, Kerpener Str. 34, 50931 Cologne, Germany. v.isachenko@yahoo.com
authors: Vladimir
authors: V
authors: Isachenko
authors: Ingrid
authors: I
authors: Orth
authors: Evgenia
authors: E
authors: Isachenko
authors: Peter
authors: P
authors: Mallmann
authors: Doris
authors: D
authors: Peters
authors: Torsten
authors: T
authors: Schmidt
authors: Bernd
authors: B
authors: Morgenstern
authors: Dolores
authors: D
authors: Foth
authors: Bettina
authors: B
authors: Hanstein
authors: Gohar
authors: G
authors: Rahimi
mesh_terms: D000818:Animals; D002645:Chickens; D049033:Chorioallantoic Membrane / Q000166:cytology; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000737:chemistry; D004958:Estradiol / Q000097:blood; D005260:Female; D005615:Freezing; D006801:Humans; D007053:Ice / Q000032:analysis; D006080:Ovarian Follicle / Q000098:blood supply / Q000000981:diagnostic imaging / Q000502:physiology / Q000637:transplantation; D010053:Ovary / Q000098:blood supply / Q000000981:diagnostic imaging / Q000502:physiology* / Q000637:transplantation*; D046509:Tissue Culture Techniques; D014182:Transplantation, Autologous; D014463:Ultrasonography; D055815:Young Adult
chemical_list: D003451:Cryoprotective Agents; D007053:Ice; D004958:Estradiol