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.
BACKGROUND: Ovarian tissue cryopreservation is the only option for preserving fertility in prepubertal girls and cancerpatients 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.
Authors: Gregory M Fahy; Brian Wowk; Jun Wu; John Phan; Chris Rasch; Alice Chang; Eric Zendejas Journal: Cryobiology Date: 2004-04 Impact factor: 2.487
Authors: J Donnez; M M Dolmans; D Demylle; P Jadoul; C Pirard; J Squifflet; B Martinez-Madrid; A van Langendonckt Journal: Lancet Date: 2004 Oct 16-22 Impact factor: 79.321
Authors: David Tagler; Tao Tu; Rachel M Smith; Nicholas R Anderson; Candace M Tingen; Teresa K Woodruff; Lonnie D Shea Journal: Tissue Eng Part A Date: 2012-03-02 Impact factor: 3.845
Authors: Monica M Laronda; Kelly E McKinnon; Alison Y Ting; Ann V Le Fever; Mary B Zelinski; Teresa K Woodruff Journal: J Assist Reprod Genet Date: 2016-11-30 Impact factor: 3.412
Authors: Jing Xu; Min Xu; Marcelo P Bernuci; Thomas E Fisher; Lonnie D Shea; Teresa K Woodruff; Mary B Zelinski; Richard L Stouffer Journal: Adv Exp Med Biol Date: 2013 Impact factor: 2.622