Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
10928096 | Cryobiology | 2013 | 6 Pages |
Abstract
To achieve optimal and uniform outcomes, slow cooling protocols for human ovarian tissues generally initiate ice formation at high sub-zero temperatures (â6 to â9 °C). The aim of the study was to investigate the function of ovarian tissue that had unintentionally self seeded at â20 °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 0.15 M sucrose which had self-seeded at â20 °C. Five years after cryopreservation, 8 pieces were thawed and transplanted back to the patient. Two small (1 Ã 2 Ã 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.
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Authors
Vladimir Isachenko, Ingrid Orth, Evgenia Isachenko, Peter Mallmann, Doris Peters, Torsten Schmidt, Bernd Morgenstern, Dolores Foth, Bettina Hanstein, Gohar Rahimi,