Clin Exp Reprod Med.  2017 Mar;44(1):8-14. 10.5653/cerm.2017.44.1.8.

Effects of supplementation with antifreeze proteins on the follicular integrity of vitrified-warmed mouse ovaries: Comparison of two types of antifreeze proteins alone and in combination

• Kim MK
• Kong HS
• Youm HW
• Jee BC

• KMID: 2408581
• DOI: http://doi.org/10.5653/cerm.2017.44.1.8

Abstract

OBJECTIVE
The aim of this study was to analyze the effect of supplementing vitrification and warming solutions with two types of antifreeze proteins (AFPs) and the combination thereof on the follicular integrity of vitrified-warmed mouse ovaries.
METHODS
Ovaries (n=154) were obtained from 5-week-old BDF1 female mice (n=77) and vitrified using ethylene glycol and dimethyl sulfoxide with the supplementation of 10 mg/mL of Flavobacterium frigoris ice-binding protein (FfIBP), 10 mg/mL of type III AFP, or the combination thereof. Ovarian sections were examined by light microscopy after hematoxylin and eosin staining, and follicular intactness was assessed as a whole and according to the type of follicle. Apoptosis within the follicles as a whole was detected by a terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick-end labeling assay.
RESULTS
The proportion of overall intact follicles was significantly higher in the type III AFP-supplemented group (60.5%) and the combination group (62.9%) than in the non-supplemented controls (43.8%, p<0.05 for each). The proportion of intact primordial follicles was significantly higher in the FfIBP-supplemented (90.0%), type III AFP-supplemented (92.3%), and combination (89.7%) groups than in the non-supplemented control group (46.2%, p<0.05 for each). The proportions of non-apoptotic follicles were similar across the four groups.
CONCLUSION
Supplementation of the vitrification and warming solutions with FfIBP, type III AFP, or the combination thereof was equally beneficial for the preservation of primordial follicles in vitrified mouse ovaries.

Keyword

Antifreeze protein; Fertility preservation; Ovary; Vitrification

MeSH Terms

Animals
Antifreeze Proteins*
Apoptosis
Deoxyuridine
Dimethyl Sulfoxide
DNA Nucleotidylexotransferase
Eosine Yellowish-(YS)
Ethylene Glycol
Female
Fertility Preservation
Flavobacterium
Hematoxylin
Humans
Mice*
Microscopy
Ovary*
Vitrification
Antifreeze Proteins
DNA Nucleotidylexotransferase
Deoxyuridine
Dimethyl Sulfoxide
Eosine Yellowish-(YS)
Ethylene Glycol
Hematoxylin