Universität zu Köln

Isachenko, Vladimir, Morgenstern, Bernd, Todorov, Plamen, Isachenko, Evgenia, Mallmann, Peter, Hanstein, Bettina and Rahimi, Gohar (2020). Long-term (24h) cooling of ovarian fragments in the presence of permeable cryoprotectants prior to freezing: Two unsuccesful IVF-cycles and spontaneous pregnancy with baby born after re-transplantation. Cryobiology, 93. S. 115 - 121. SAN DIEGO: ACADEMIC PRESS INC ELSEVIER SCIENCE. ISSN 1090-2392

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Abstract

Cancer is the second major cause of death in the world. The problem of post-cancer infertility plays a significant role, because chemotherapy can be gonadotoxic. Cryopreservation of ovarian tissue before cancer therapy with re-implantation after convalescence is the potential key solution to this problem. The aim of this study was to test the viability of cryopreserved human ovarian cortex after long-term cooling in culture medium composed of permeable cryoprotectants. Ovarian fragments from sixteen patients were randomly divided into two groups. After the operation, tissue pieces assigned to both groups were cooled to 5 degrees C for 22-24 h, frozen and thawed. Group 1 pieces (n = 32) were cooled before cryopreservation in the standard culture medium, and Group 2 pieces (n = 32) were cooled in the freezing medium (culture medium+6% ethylene glycol+6% dimethyl sulfoxide+0.15 M sucrose). Freezing was performed in standard 5 ml cryo-vials with ice formation at -9 degrees C, cooling from -9 to -34 degrees C at a rate of -0.3 degrees C/min and plunging at -34 degrees C into liquid nitrogen. After thawing in a 100 degrees C (boiling) water bath, the removal of cryoprotectants was performed in 0.5 M sucrose with 20 min exposure in sucrose and 30 min stepping rehydration. The effectiveness of the pre-freezing cooling of tissue was evaluated by the development of follicles (histology). Six months after the autotransplantation, oocytes from the twenty-seven-year old, hormonally stimulated patient were retrieved and fertilized with her partner sperm through the intracytoplasmic spermatozoa injection (ICSI). For groups 1 and 2, 93.5 +/- 1.9% and 96.4 +/- 2.0% of the preantral follicles, respectively, were morphologically normal (P > 0.1) (with a tendency toward increasing in quality in Group 2). Six months after the auto-transplantation, two ICSI cycles resulted in the gathering and transplantation of high quality embryos, but no pregnancy had been established. Thirteen months after the autotransplantation, the patient became spontaneously pregnant and delivered a healthy baby girl at term. Long-term (24 h) cooling of ovarian tissue to 5 degrees C before cryopreservation in the presence of permeable cryoprotectants simplifies the protocol of cryopreservation and has a tendency of increasing of the cells viability after thawing.

Item Type:	Journal Article
Creators:	Creators Email ORCID ORCID Put Code Isachenko, Vladimir UNSPECIFIED UNSPECIFIED UNSPECIFIED Morgenstern, Bernd UNSPECIFIED UNSPECIFIED UNSPECIFIED Todorov, Plamen UNSPECIFIED UNSPECIFIED UNSPECIFIED Isachenko, Evgenia UNSPECIFIED UNSPECIFIED UNSPECIFIED Mallmann, Peter UNSPECIFIED UNSPECIFIED UNSPECIFIED Hanstein, Bettina UNSPECIFIED UNSPECIFIED UNSPECIFIED Rahimi, Gohar UNSPECIFIED UNSPECIFIED UNSPECIFIED
URN:	urn:nbn:de:hbz:38-338731
DOI:	10.1016/j.cryobiol.2020.01.022
Journal or Publication Title:	Cryobiology
Volume:	93
Page Range:	S. 115 - 121
Date:	2020
Publisher:	ACADEMIC PRESS INC ELSEVIER SCIENCE
Place of Publication:	SAN DIEGO
ISSN:	1090-2392
Language:	English
Faculty:	Unspecified
Divisions:	Unspecified
Subjects:	no entry
Uncontrolled Keywords:	Keywords Language IN-VITRO; TISSUE; CRYOPRESERVATION; FERTILIZATION; SPERMATOZOA Multiple languages Biology; Physiology Multiple languages
URI:	http://kups.ub.uni-koeln.de/id/eprint/33873