Clin Exp Reprod Med.  2018 Sep;45(3):143-148. 10.5653/cerm.2018.45.3.143.

Angiopoietin-1 and -2 and vascular endothelial growth factor expression in ovarian grafts after cryopreservation using two methods

• Cho IA
• Lee YJ
• Lee HJ
• Choi IY
• Shin JK
• Lee SA
• Lee JH
• Choi WJ

• KMID: 2419531
• DOI: http://doi.org/10.5653/cerm.2018.45.3.143

Abstract

OBJECTIVE
The favored method of preserving fertility in young female cancer survivors is cryopreservation and autotransplantation of ovarian tissue. Reducing hypoxia until angiogenesis takes place is essential for the survival of transplanted ovarian tissue. The aim of this study was to investigate the role of angiopoietin-1 (Angpt-1), angiopoietin-2 (Angpt-2), and vascular endothelial growth factor (VEGF) in ovarian tissue grafts that were cryopreserved using two methods.
METHODS
Ovarian tissues harvested from ICR mice were divided into three groups: group I (control), no cryopreservation; group II, vitrification in EFS (ethylene-glycol, ficoll, and sucrose solution)-40; and group III, slow freezing in dimethyl sulfoxide. We extracted mRNA for VEGF, Angpt-1, and Angpt-2 from ovarian tissue 1 week following cryopreservation and again 2 weeks after autotransplantation. We used reverse transcriptase-polymerase chain reaction to quantify the levels of VEGF, Angpt-1, and Angpt-2 in the tissue.
RESULTS
Angpt-1 and Angpt-2 expression decreased after cryopreservation in groups II and III. After autotransplantation, Angpt-1 and Angpt-2 expression in ovarian tissue showed different trends. Angpt-1 expression in groups II and III was lower than in group I, but Angpt-2 in groups II and III showed no significant difference from group I. The vitrified ovarian tissues had higher expression of VEGF and Angpt-2 than the slowfrozen ovarian tissues, but the difference was not statistically significant.
CONCLUSION
Our results indicate that Angpt-2 may play an important role in ovarian tissue transplantation after cryopreservation although further studies are needed to understand its exact function.

Keyword

Angiopoietin-1; Angiopoietin-2; Autografts; Cryopreservation; Fertility preservation; Mice; Neovascularization; Ovary; Reverse transcriptase polymerase chain reaction; Vascular endothelial growth factors

MeSH Terms

Angiopoietin-1*
Angiopoietin-2
Animals
Anoxia
Autografts
Cryopreservation*
Dimethyl Sulfoxide
Female
Fertility
Fertility Preservation
Ficoll
Freezing
Humans
Methods*
Mice
Mice, Inbred ICR
Ovary
Reverse Transcriptase Polymerase Chain Reaction
RNA, Messenger
Sucrose
Survivors
Tissue Transplantation
Transplantation, Autologous
Transplants*
Vascular Endothelial Growth Factor A*
Vascular Endothelial Growth Factors
Vitrification
Angiopoietin-1
Angiopoietin-2
Dimethyl Sulfoxide
Ficoll
RNA, Messenger
Sucrose
Vascular Endothelial Growth Factor A
Vascular Endothelial Growth Factors