Viability of human ovarian tissue confirmed 5 years after freezing with spontaneous ice-formation by autografting and chorio-allantoic membrane culture

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• 作者：Vladimir Isachenko ; ^{v.isachenko@yahoo.com} ; Ingrid Orth ; Evgenia Isachenko ; Peter Mallmann ; Doris Peters ; Torsten Schmidt ; Bernd Morgenstern ; Dolores Foth ; Bettina Hanstein ; Gohar Rahimi
• 关键词：Human ; Ovarian tissue ; Freezing ; Ice ; Transplantation ; Follicles ; Angiogenesis
• 刊名：Cryobiology
• 出版年：2013
• 出版时间：June, 2013
• 年：2013
• 卷：66
• 期：3
• 页码：233-238
• 全文大小：946 K

文摘

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 ¡ãC). The aim of the study was to investigate the function of ovarian tissue that had unintentionally self seeded at ?20 ¡ãC 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 ?20 ¡ãC. Five years after cryopreservation, 8 pieces were thawed and transplanted back to the patient. Two small (1 ¡Á 2 ¡Á 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-¦Â 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.