Effect of necrostatin on mouse ovarian cryopreservation and transplantation

Jung Ryeol Lee; Hye Won Youm; Seul Ki Kim; Byung Chul Jee; Chang Suk Suh; Seok Hyun Kim

doi:10.1016/j.ejogrb.2014.04.040

Effect of necrostatin on mouse ovarian cryopreservation and transplantation

Eur J Obstet Gynecol Reprod Biol. 2014 Jul:178:16-20. doi: 10.1016/j.ejogrb.2014.04.040. Epub 2014 May 13.

Authors

Jung Ryeol Lee¹, Hye Won Youm², Seul Ki Kim³, Byung Chul Jee², Chang Suk Suh², Seok Hyun Kim⁴

Affiliations

¹ 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.
² 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.
³ Department of Obstetrics and Gynaecology, Seoul National University Bundang Hospital, Seongnam, Republic of Korea.
⁴ Department of Obstetrics and Gynaecology, Seoul National University College of Medicine, Seoul, Republic of Korea. Electronic address: seokhyun@snu.ac.kr.

PMID: 24948049
DOI: 10.1016/j.ejogrb.2014.04.040

Abstract

Objective: To investigate the effects of necrostatin-1 (Nec-1) supplementation on vitrification, warming and transplantation of ovarian tissue.

Study design: 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.

Keywords: Fertility preservation; Necrostatin; Ovarian tissue cryopreservation; Vitrification.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Apoptosis / drug effects
Cryopreservation / methods*
Female
Fertility Preservation
Imidazoles / pharmacology*
In Situ Nick-End Labeling
Indoles / pharmacology*
Mice, Inbred ICR
Ovarian Follicle / drug effects*
Ovary / drug effects*
Ovary / transplantation*
Vitrification

Substances

Imidazoles
Indoles
necrostatin-1