Cryopreservation of human ovarian tissue using the silver closed vitrification system
The aim of this study is to evaluate the feasibility of using a hand-made silver container for the cryopreservation of human ovarian cortex.
Human ovarian cortex tissues were vitrified using an open vitrification system (OVS) of needle immersed vitrification (NIV) and two closed vitrification systems (CVS) of a plastic vial (plastic CVS) and a silver container (silver CVS). Outcomes of vitrification were evaluated morphologically and histologically by in vitro culture and xenotransplantation. The apoptosis of primordial follicles was assessed by TUNEL staining. The production of E2 and P4 was examined by a chemiluminescent immunoassay. Blood vessels were visualized with CD31 staining.
Compared with the fresh ovarian cortex tissue, ovarian cortex tissues that were vitrified using the three different carriers and then warmed showed significantly reduced percentages of normal primordial follicles, viability of primordial follicles, E2 and P4 levels during in vitro culture and decreased amounts of blood vessels. However, much better outcomes were obtained with NIV and silver CVS than with plastic CVS, based on the better morphology and viability of primordial follicles, higher E2 and P4 production during an in vitro culture, and greater numbers of blood vessels after xenografting. Importantly, the outcomes of ovarian cortex cryopreservation with silver CVS were similar and comparable to those with NIV.
The hand-made silver container as a CVS is a promising carrier for the cryopreservation of the human ovarian cortex.
KeywordsClosed vitrification system Silver container Ovarian cortex Primordial follicles Needle immersed vitrification
This research was financially supported by Science and Technology Bureau of Chengdu City (2014-HM01-00045-SF). All the authors are very grateful for the technical support from Dr. Yan Wang from Stanford University and Dr. Yong-Can Huang from the University of Hong Kong for manuscript revision.
Zhun Xiao design of the study, revising it critically for important intellectual content; Yaoyao Zhang acquisition of data, drafting the article; Wei Fan analysis and interpretation of data, all authors approve the version to be submitted.
Compliance with ethical standards
Conflicts of interest
The authors declare that they have no conflict of interest.
- 4.Jensen AK, Macklon KT, Fedder J, Ernst E, Humaidan P, Anderson CY, et al. 86 successful births and 9 ongoing pregnancies worldwide in women transplanted with frozen-thawed ovarian tissue:focus on birth and perinatal outcome in 40 of these children. J Assist Reprod Genet. 2017;34:325–36.CrossRefPubMedGoogle Scholar
- 7.Donnez J, Dolmans MM, Demylle D, Jadoul P, Pirard C, Squifflet J, et al. Restoration of ovarian function after orthotopic (intraovarian and periovarian) transplantation of cryopreserved ovarian tissue in a woman treated by bone marrow transplantation for sickle cell anaemia: case report. Hum Reprod. 2006;211:183–8.CrossRefGoogle Scholar
- 18.Lin TC, Yen JM, Kuo TC, Gong KB, Hsu KH, Hsu TT. Comparison of the developmental potential of 2-week-old preantral follicles derived from vitrified ovarian tissue slices, vitrified whole ovaries and vitrified/transplanted newborn mouse ovaries using the metal surface method. BMC Biotechnol. 2008;8:38.CrossRefPubMedPubMedCentralGoogle Scholar
- 26.Bos-Mikich A, Marques L, Rodrigues JL, Lothhammer N, Frantz N. The use of a metal container for vitrification of mouse ovaries, as a clinical grade model for human ovarian tissue cryopreservation, after different times and temperatures of transport. J Assist Reprod Genet. 2012;2911:1267–71.CrossRefGoogle Scholar
- 28.Makinson R. The thermal conductivity of metals. In: mathematical proceedings of the Cambridge philosophical society. Cambridge: Univ Press; 1938. p. 474–97.Google Scholar
- 40.Abir R, Fisch B, Fisher N, Samara N, Lerer-Serfaty G, Magen R, et al. Attempts to improve human ovarian transplantation outcomes of needle-immersed vitrification and slow-freezing by host and graft treatments. J Assist Reprod Genet. 2017;34:633–44. doi: 10.1007/s10815-017-0884-8.CrossRefPubMedGoogle Scholar
- 41.Younis AJ, Lerer-Serfaty G, Stav D, Sabbah B, Shochat T, Kessler-Icekson G, et al. Extracellular-like matrices and leukaemia inhibitory factor for in vitro culture of human primordial follicles. Reprod Fertil Dev. 2017; doi: 10.1071/RD16233.
- 45.Simopoulou M, Asimakopoulos B, Bakas P, Boyadjiev N, Tzanakaki D, Creatsas G. Oocyte and embryo vitrification in the IVF laboratory: a comprehensive review. Folia Med (Plovdiv). 2014;563:161–9.Google Scholar