In vitro development of secondary follicles from cryopreserved rhesus macaque ovarian tissue after slow-rate freeze or vitrification

Alison Ting, Richard R. Yeoman, Maralee S. Lawson, Mary Zelinski

Research output: Contribution to journalArticle

77 Citations (Scopus)

Abstract

Background Ovarian tissue cryopreservation is the only option for preserving fertility in prepubertal girls and cancer patients requiring immediate treatment. Following ovarian tissue cryopreservation, fertility can be restored after tissue transplant or in vitro follicle maturation. Methods Macaque (n = 4) ovarian cortex was cryopreserved using slow-rate freezing (slow freezing) or vitrification. Tissues were fixed for histology or phosphohistone H3 (PPH3) analysis, cultured with bromodeoxyuridine (BrdU) or used for three-dimensional secondary follicle culture. Follicular diameter and steroid hormones were measured weekly. Results Slow freezing induced frequent cryo-injuries while vitrification consistently maintained morphology of the stroma and secondary follicles. PPH3 was similar in fresh and vitrified, but sparse in slow-frozen tissues. BrdU uptake appeared diminished following both Methods compared with that in fresh follicles. In vitro follicle survival and growth were greater in fresh than in cryopreserved follicles. Antrum formation appeared similar after vitrification compared with the fresh, but was reduced following slow freezing. Steroid production was delayed or diminished following both Methods compared with fresh samples. Conclusions Secondary follicle morphology was improved after vitrification relative to slow freezing. Following vitrification, stroma was consistently more compact with intact cells typical to that of fresh tissue. BrdU uptake demonstrated follicle viability post-thaw/warming. For the first time, although not to the extent of fresh follicles, macaque follicles from cryopreserved tissue can survive, grow, form an antrum and produce steroid hormones, indicating some functional preservation. The combination of successful ovarian tissue cryopreservation with in vitro maturation of follicles will offer a major advancement to the field of fertility preservation.

Original languageEnglish (US)
Pages (from-to)2461-2472
Number of pages12
JournalHuman Reproduction
Volume26
Issue number9
DOIs
StatePublished - Sep 2011

Fingerprint

Vitrification
Macaca mulatta
Freezing
Cryopreservation
Bromodeoxyuridine
Steroids
Macaca
Fertility
Hormones
Fertility Preservation
In Vitro Techniques
Histology
Transplants
Survival
Wounds and Injuries
Growth

Keywords

  • Cryopreservation
  • in vitro follicle culture
  • non-human primates
  • ovarian tissue
  • vitrification

ASJC Scopus subject areas

  • Rehabilitation
  • Obstetrics and Gynecology
  • Reproductive Medicine

Cite this

In vitro development of secondary follicles from cryopreserved rhesus macaque ovarian tissue after slow-rate freeze or vitrification. / Ting, Alison; Yeoman, Richard R.; Lawson, Maralee S.; Zelinski, Mary.

In: Human Reproduction, Vol. 26, No. 9, 09.2011, p. 2461-2472.

Research output: Contribution to journalArticle

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