Journal of Assisted Reproduction and Genetics

, Volume 34, Issue 11, pp 1435–1444 | Cite as

Cryopreservation of human ovarian tissue using the silver closed vitrification system

  • Zhun XiaoEmail author
  • Yaoyao Zhang
  • Wei Fan
Fertility Preservation



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.


Closed 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.

Author contributions

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.


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Copyright information

© Springer Science+Business Media, LLC 2017

Authors and Affiliations

  1. 1.Reproductive Medical Center of West China 2nd Hospital, Key Laboratory of Birth Defects and Related Diseases of Women and Children, Ministry of EducationSichuan UniversityChengduP. R. China

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