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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

The electron micrograph of granulosa cells of a secondary follicle. A and B) vitrified ovaries in single cryoprotectant, C) vitrified ovaries in double cryoprotectant, showing arrangement of mithochondria m, numerous lipid droplets Lp, vacuole V and atypical or irregularly shaped mithochondria with longitudinally oriented cristae were found in CV1-4 groups or single cryoprotectant, TEM: ×10000. B; numerous spherical mithochondria with continuous membranes and normal cristae were obserced in CV6, 7 groups or double cryoprotectant, A; TEM: ×4646, B and C; TEM: ×10000.
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Fig4: The electron micrograph of granulosa cells of a secondary follicle. A and B) vitrified ovaries in single cryoprotectant, C) vitrified ovaries in double cryoprotectant, showing arrangement of mithochondria m, numerous lipid droplets Lp, vacuole V and atypical or irregularly shaped mithochondria with longitudinally oriented cristae were found in CV1-4 groups or single cryoprotectant, TEM: ×10000. B; numerous spherical mithochondria with continuous membranes and normal cristae were obserced in CV6, 7 groups or double cryoprotectant, A; TEM: ×4646, B and C; TEM: ×10000.

Mentions: The ultrastractural analysis revealed that secondary follicles of ovaries frozen in CV7 group had morphologically normal similar to those preserves in non vitrified (control) ovaries and the integrity of cell organelles was well preserved (Figure 3B and C, Figure 4C).Figure 4


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)

The electron micrograph of granulosa cells of a secondary follicle. A and B) vitrified ovaries in single cryoprotectant, C) vitrified ovaries in double cryoprotectant, showing arrangement of mithochondria m, numerous lipid droplets Lp, vacuole V and atypical or irregularly shaped mithochondria with longitudinally oriented cristae were found in CV1-4 groups or single cryoprotectant, TEM: ×10000. B; numerous spherical mithochondria with continuous membranes and normal cristae were obserced in CV6, 7 groups or double cryoprotectant, A; TEM: ×4646, B and C; TEM: ×10000.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

Fig4: The electron micrograph of granulosa cells of a secondary follicle. A and B) vitrified ovaries in single cryoprotectant, C) vitrified ovaries in double cryoprotectant, showing arrangement of mithochondria m, numerous lipid droplets Lp, vacuole V and atypical or irregularly shaped mithochondria with longitudinally oriented cristae were found in CV1-4 groups or single cryoprotectant, TEM: ×10000. B; numerous spherical mithochondria with continuous membranes and normal cristae were obserced in CV6, 7 groups or double cryoprotectant, A; TEM: ×4646, B and C; TEM: ×10000.
Mentions: The ultrastractural analysis revealed that secondary follicles of ovaries frozen in CV7 group had morphologically normal similar to those preserves in non vitrified (control) ovaries and the integrity of cell organelles was well preserved (Figure 3B and C, Figure 4C).Figure 4

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