title: Effect of necrostatin on mouse ovarian cryopreservation and transplantation.
abstract: OBJECTIVE
  To investigate the effects of necrostatin-1 (Nec-1) supplementation on vitrification, warming and transplantation of ovarian tissue.
  
  
  METHODS
  Ovaries from 4-week-old ICR mice were vitrified using a two-step procedure; ovaries were suspended in equilibration solution for 10min, and then mixed with vitrification solution for 5min. Ovaries were divided at random into three groups and 0 (control), 25 or 100μM Nec-1 was added to the vitrification solution. After warming, follicular morphology and apoptosis were assessed. For each group, a sample of vitrified, warmed ovaries was autotransplanted. The same dose of Nec-1 that was added to the vitrification solution was added to each warming solution and injected intraperitoneally. Follicular morphology and apoptosis of transplanted ovaries were assessed after 2 weeks.
  
  
  RESULTS
  After vitrification and warming, morphological analysis revealed that the intact follicle ratio was significantly higher in the Nec-1-treated groups compared with the control group (control, 45.1%; 25μM Nec-1, 51.7%; 100μM Nec-1, 57.9%). The rate of apoptosis was lower in the Nec-1 treated groups compared with the control group (control, 11.2%; 25μM Nec-1, 8.5%; 100μM Nec-1, 7.2%). After transplantation of the vitrified, warmed ovaries, morphological analysis revealed that the intact follicle ratio was significantly higher in the Nec-1 treated groups compared with the control group (control, 43.1%; 25μM Nec-1, 60.6%; 100μM Nec-1, 70.7%). The rate of apoptosis was lower in the Nec-1 treated groups compared with the control group (control, 5.3%; 25μM Nec-1, 2.5%; 100μM Nec-1, 2.0%).
  
  
  CONCLUSIONS
  Nec-1 supplementation during vitrification, warming and transplantation has beneficial effects on the survival of ovarian tissue. These results can help to improve ovarian tissue vitrification and transplantation protocols for fertility preservation.
authors.affiliation: Department of Obstetrics and Gynaecology, Seoul National University Bundang Hospital, Seongnam, Republic of Korea; Department of Obstetrics and Gynaecology, Seoul National University College of Medicine, Seoul, Republic of Korea. Electronic address: leejrmd@snu.ac.kr.
authors.forename: Jung Ryeol
authors.initials: JR
authors.lastname: Lee
authors.affiliation: Department of Obstetrics and Gynaecology, Seoul National University Bundang Hospital, Seongnam, Republic of Korea; Department of Obstetrics and Gynaecology, Seoul National University College of Medicine, Seoul, Republic of Korea.
authors.forename: Hye Won
authors.initials: HW
authors.lastname: Youm
authors.affiliation: Department of Obstetrics and Gynaecology, Seoul National University Bundang Hospital, Seongnam, Republic of Korea.
authors.forename: Seul Ki
authors.initials: SK
authors.lastname: Kim
authors.affiliation: Department of Obstetrics and Gynaecology, Seoul National University Bundang Hospital, Seongnam, Republic of Korea; Department of Obstetrics and Gynaecology, Seoul National University College of Medicine, Seoul, Republic of Korea.
authors.forename: Byung Chul
authors.initials: BC
authors.lastname: Jee
authors.affiliation: Department of Obstetrics and Gynaecology, Seoul National University Bundang Hospital, Seongnam, Republic of Korea; Department of Obstetrics and Gynaecology, Seoul National University College of Medicine, Seoul, Republic of Korea.
authors.forename: Chang Suk
authors.initials: CS
authors.lastname: Suh
authors.affiliation: Department of Obstetrics and Gynaecology, Seoul National University College of Medicine, Seoul, Republic of Korea. Electronic address: seokhyun@snu.ac.kr.
authors.forename: Seok Hyun
authors.initials: SH
authors.lastname: Kim
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods*; D005260:Female; D059247:Fertility Preservation; D007093:Imidazoles / Q000494:pharmacology*; D020287:In Situ Nick-End Labeling; D007211:Indoles / Q000494:pharmacology*; D008813:Mice, Inbred ICR; D006080:Ovarian Follicle / Q000187:drug effects*; D010053:Ovary / Q000187:drug effects* / Q000637:transplantation*; D058989:Vitrification
chemical_list: D007093:Imidazoles; D007211:Indoles; C507699:necrostatin-1
keywords: Fertility preservation; Necrostatin; Ovarian tissue cryopreservation; Vitrification