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J Ovarian Res. 2015 Mar 6;8:7. doi: 10.1186/s13048-015-0137-3.

Preservation of mouse ovarian tissue follicle morphology and ultra-structure after vitrifying in biotechnological protocols.

Abstract

BACKGROUND:

The aim of the present study was to characterize the morphological and ultrastractural of mouse ovarian tissue with different cryoprotectant solution.

OBJECTIVE:

Aim of this study, is to demonstrae an improved convetional vitrification method on mouse ovarian tissue using different concentrations of ethylene glycol (EG) and/or dimetyl sulfoxide (DMSO) and EG.

MATERIALS AND METHODS:

Mouse ovarian tissue dissected and were randomly assigned to three groups: control, conventional vitrification (CV) and toxicity test. Then ovaries were vitrified by 5%, 10% EG or DMSO CV1-CV4, 5%, 10% EG plus DMSO CV5-CV6 and EG plus DMSO in climbing concentrations CV7. The effect of cryoprotectant solutions on ovarian tissue were evaluated by histological examination hematotoxillin & eosin stain, H&E, viability assessment trypan blue stain and ultrastructural analyses transmission electron microscopy, TEM.

RESULTS:

Ovarian tissue frozen in CV7 solution showed a higher percentage of morphologically normal follicles or viable follicles than other cryoprotectant solutions P < 0.05. Ultrastructural analysis of ovarian tissue showed that less damage was observed in CV7 and it was very similar to the control group.

CONCLUSION:

Vitrification of ovarian tissue with optimal cryoprotectant solutions such as EG plus DMSO is the most effective for preserving the structural efficiency of ovarian follicles.

PMID:
25824613
PMCID:
PMC4356062
DOI:
10.1186/s13048-015-0137-3
[Indexed for MEDLINE]
Free PMC Article

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