Morphologic, ultrastructural, and biochemical identification of apoptosis in vitrified-warmed mouse ovarian tissue

Mazoochi, Tahere; Salehnia, Mojdeh; Valojerdi, Mojtaba Rezazadeh; Mowla, Seyed Javad

doi:10.1016/j.fertnstert.2007.07.1384

Cited by 38 publications

(25 citation statements)

References 31 publications

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“…In follicles included in ovarian tissue that were cryopreserved in the presence of EG, greater damage was observed in this study, manifested as reduced electron density and fewer organelles in the ooplasm. These findings differed from those in goats [2] and cattle [31], in which the cryoprotectant agent was highly efficient in maintaining follicle ultrastructure.…”

Section: Discussioncontrasting

confidence: 87%

Ultrastructural features of agouti (Dasyprocta aguti) preantral follicles cryopreserved using dimethyl sulfoxide, ethylene glycol and propanediol

Wanderley¹,

Luz²,

Faustino³

et al. 2012

Theriogenology

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].You may download, copy and otherwise use the AAM for non-commercial purposes provided that your license is limited by the following restrictions:(1) You may use this AAM for non-commercial purposes only under the terms of the CC-BY-NC-ND license.(2) The integrity of the work and identification of the author, copyright owner, and publisher must be preserved in any copy. Ultrastructural features of agouti (Dasyprocta aguti) preantral follicles cryopreserved using dimethyl sulfoxide, ethylene glycol and propanediol AbstractThe objective was to develop an efficient protocol for cryopreservation of agouti (Dasyprocta aguti) ovarian tissue. Agouti ovarian fragments were placed, for 10 min, in a solution containing MEM and fetal bovine serum plus 1.5 M dimethyl sulfoxide (DMSO), ethylene glycol (EG) or propanediol (PROH); some of those fragments were subsequently cryopreserved in a programmable freezer. After exposure and/or thawing, all samples were fixed in Carnoy prior to histological analysis. To evaluate ultrastructure, follicles from the control and all cryopreserved treatments were fixed in Karnovsky and processed for transmission electron microscopy. After exposure and freezing, there was a significant decrease in the percentage of morphologically normal preantral follicles in all treatments when compared to the control (92.67 ± 2.79, mean ± SD). However, there were no significant difference when the exposure and freezing procedures were compared using the same cryoprotectant. Moreover, there was no significant difference among cryoprotectants at the time of exposure (DMSO: 64.7 ± 3.8; EG: 70.7 ± 11.2, PROH: 63.3 ± 8.5) or after freezing (DMSO: 60.6 ± 3.6, EG: 64.0 ± 11.9; PROH: 62.0 ± 6.9). However, only follicles frozen with PROH had normal ultrastructure. In conclusion, preantral follicles enclosed in agouti ovarian tissue were successfully cryopreserved using 1.5 M PROH, with satisfactory maintenance of follicle morphology and ultrastructure.

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Section: Discussioncontrasting

confidence: 87%

Ultrastructural features of agouti (Dasyprocta aguti) preantral follicles cryopreserved using dimethyl sulfoxide, ethylene glycol and propanediol

Wanderley¹,

Luz²,

Faustino³

et al. 2012

Theriogenology

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“…In contrast, other investigations and our previous report have shown that the incidence of apoptosis in cryopreserved tissue was not significantly different from fresh controls (Demirci et al, 2002;Bedaiwy and Hossein, 2004;Hussein et al, 2006;Mazoochi et al, 2008).…”

Section: Introductioncontrasting

confidence: 91%

“…There are controversial reports about the effects of cryopreservation on the incidence of apoptosis in ovarian tissue after thawing (Demirci et al, 2002;Bedaiwy and Hossein, 2004;Rimon et al, 2005;Tirelli et al, 2005;Hussein et al, 2006;Mazoochi et al, 2008). Tirelli et al (2005) observed a significant increase in apoptosis of sow granulosa cells that were frozen slowly.…”

Section: Introductionmentioning

confidence: 99%

Analysis of apoptosis and expression of genes related to apoptosis in cultures of follicles derived from vitrified and non-vitrified ovaries

Mazoochi

Salehnia

Pourbeiranvand

et al. 2009

Molecular Human Reproduction

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The aim of this study was to evaluate the incidence of apoptosis after in vitro culture of isolated follicles derived from vitrified and non-vitrified ovaries. Mouse ovaries were vitrified and their pre-antral follicles were mechanically isolated and cultured for 10 days. Growth and survival rates of the follicles were assessed during the culture period and the ultrastructure of the follicles was studied. The expression of p53, Bcl-2, Bax, Fas, FasL and survivin were analyzed by real-time RT -PCR in different follicular developmental stages. The percentages of apoptotic and necrotic cells were determined using a fluorescein-activated cell sorting (FACS) technique. There were no differences between the growth and survival rates of follicles in the vitrified and non-vitrified groups. All of the evaluated genes were expressed in the pre-antral, large pre-antral and antral follicles in both groups, except Fas mRNA, which was not expressed in the pre-antral follicles. The expression of p53, Bcl2, Bax and FasL mRNA was similar in vitrified and non-vitrified groups; however, Fas mRNAs were more strongly expressed in the antral follicles of the vitrified group than of the control group (P , 0.05). The expression of survivin 140 was lower in the antral follicles of the vitrified group than of the control group (P , 0.05). FACS analysis showed that the percentage of intact cells was lower in the vitrified group than in the non-vitrified group (P , 0.05). This study demonstrated no signs of apoptosis ultrastructurally in cultured follicles; however, vitrification was shown to affect the expression of some genes related to apoptosis.

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“…They concluded that cryopreservation caused the death of ovarian cells through apoptosis and thus inhibiting further development of primordial follicles [61]. However a study by Mazoochi et al(2008) [62] carried out in the mouse showed that there was no sign of apoptosis in vitrified and cultured follicles [62]. A study carried out with human ovarian tissue also reported that vitrification has no effect on apoptosis of follicles [63,64].…”

Section: Discussionmentioning

confidence: 99%

“…During vitrification fluid in the cell is converted to the solid state, avoiding cellular damage caused by the formation of intracellular ice crystals [6]. Vitrification also does not induce apoptosis in ovarian tissue after warming [7,8]. Recent studies reported that the use of vitrification yielded higher oocyte survival rates compared to slow freezing [4,9].…”

Section: Introductionmentioning

confidence: 99%

In vitro development of human primordial follicles to preantral stage after vitrification

Khosravi

Reid

Moini

et al. 2013

J Assist Reprod Genet

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Purpose The aim was to culture primordial follicles in vitro to reach preantral stage in vitrified human ovarian tissue. Methods Ovarian tissue samples were obtained from six women. Tissue strips were vitrified by infiltration with a cryoprotectant followed by mounting on a stainless steel carrier. After culturing for 7 days the morphology and developmental stages of follicles enclosed in fresh and vitrified groups were analyzed. Results High proportion of viable follicles in vitrified ovarian strips was obtained. After culturing for 7 days the percentage of secondary and preantral follicles increased significantly (P <0.05) whereas primordial and transitory follicles showed a significant decrease (P <0.05) compared to their respective counterparts at day 0 of culture.Conclusions Vitrification of ovarian strips with an improved carrier device and culturing of follicles in ovarian strips after warming yielded developed follicles with high viability and morphological integrity that may be suitable for use in fertility preservation among cancer patients.

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Morphologic, ultrastructural, and biochemical identification of apoptosis in vitrified-warmed mouse ovarian tissue

Cited by 38 publications

References 31 publications

Ultrastructural features of agouti (Dasyprocta aguti) preantral follicles cryopreserved using dimethyl sulfoxide, ethylene glycol and propanediol

Ultrastructural features of agouti (Dasyprocta aguti) preantral follicles cryopreserved using dimethyl sulfoxide, ethylene glycol and propanediol

Analysis of apoptosis and expression of genes related to apoptosis in cultures of follicles derived from vitrified and non-vitrified ovaries

In vitro development of human primordial follicles to preantral stage after vitrification

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