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

Tayefi Nasrabadi H, Gavami M, Akbarzadeh A, Beheshti R, Mohammadnejad D, Abedelahi A - J Ovarian Res (2015)

Bottom Line: 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.Ovarian tissue frozen in CV7 solution showed a higher percentage of morphologically normal follicles or viable follicles than other cryoprotectant solutions P < 0.05.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.

View Article: PubMed Central - PubMed

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.

No MeSH data available.


Related in: MedlinePlus

Transmission electron microscopy images of the secondary follicle from single cryoprotectants. The zona pellucid ZP was fully developed and forming a thick layer around the oocyte O, the oocyte had swollen mitochondria m and their crista had disappeared. Many wider spaces S observed between oocyte and granulosa cells GC. A) TEM: ×1670, B) TEM: ×2784.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
getmorefigures.php?uid=PMC4356062&req=5

Fig5: Transmission electron microscopy images of the secondary follicle from single cryoprotectants. The zona pellucid ZP was fully developed and forming a thick layer around the oocyte O, the oocyte had swollen mitochondria m and their crista had disappeared. Many wider spaces S observed between oocyte and granulosa cells GC. A) TEM: ×1670, B) TEM: ×2784.

Mentions: After vitrifying/thawing in single cryoprotectant, the cytoplasm organelles were greatly changed in shape and distribution. The oocyte had most vacuole with nuclear shrinkage and elongated mitochondria with a few cristae (Figure 4A and B). In granulose cells, the mitochondria swollen and cristae disappeared. And also, these cells exhibited irregularly-shaped nuclei, irregular distribution of cytoplasmic organelles and vacuolated space (Figure 4A and B). Also wider space observed between oocyte and granulose cells or around of zona pellucida (Figure 5A and B).Figure 5


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

Tayefi Nasrabadi H, Gavami M, Akbarzadeh A, Beheshti R, Mohammadnejad D, Abedelahi A - J Ovarian Res (2015)

Transmission electron microscopy images of the secondary follicle from single cryoprotectants. The zona pellucid ZP was fully developed and forming a thick layer around the oocyte O, the oocyte had swollen mitochondria m and their crista had disappeared. Many wider spaces S observed between oocyte and granulosa cells GC. A) TEM: ×1670, B) TEM: ×2784.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4356062&req=5

Fig5: Transmission electron microscopy images of the secondary follicle from single cryoprotectants. The zona pellucid ZP was fully developed and forming a thick layer around the oocyte O, the oocyte had swollen mitochondria m and their crista had disappeared. Many wider spaces S observed between oocyte and granulosa cells GC. A) TEM: ×1670, B) TEM: ×2784.
Mentions: After vitrifying/thawing in single cryoprotectant, the cytoplasm organelles were greatly changed in shape and distribution. The oocyte had most vacuole with nuclear shrinkage and elongated mitochondria with a few cristae (Figure 4A and B). In granulose cells, the mitochondria swollen and cristae disappeared. And also, these cells exhibited irregularly-shaped nuclei, irregular distribution of cytoplasmic organelles and vacuolated space (Figure 4A and B). Also wider space observed between oocyte and granulose cells or around of zona pellucida (Figure 5A and B).Figure 5

Bottom Line: 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.Ovarian tissue frozen in CV7 solution showed a higher percentage of morphologically normal follicles or viable follicles than other cryoprotectant solutions P < 0.05.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.

View Article: PubMed Central - PubMed

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.

No MeSH data available.


Related in: MedlinePlus