The preservation of vital functions in cat ovarian tissues during vitrification depends more on the temperature of the cryoprotectant exposure than on the sucrose supplementation

Mouttham, Lara; Comizzoli, Pierre

doi:10.1016/j.cryobiol.2016.07.013

Cited by 34 publications

(34 citation statements)

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“…After freezing and thawing, numbers and percentages of MNFs detected were not significantly different from those found in fresh control samples, which is consistent with our previous findings [21]. Our results are similar to the ones reported on vitrification of cat ovarian tissue using DMSO supplemented with sucrose [18]. Using the same cryoprotectants, Tanpradit et al [16] reported better results in slow freezing than vitrification of cat ovarian tissue for follicular viability, histology, and apoptosis assessment.…”

Section: Discussionsupporting

confidence: 92%

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Function of Cryopreserved Cat Ovarian Tissue after Autotransplantation

Vilela

Leonel

Gonçalves

et al. 2019

Animals

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The aim of this study was to assess a slow-freezing protocol of cat ovarian tissue cryopreservation using autotransplantation. Four adult queens were ovariohysterectomized and the ovaries were fragmented and cryopreserved. After one week, the grafts were thawed and autografted to the subcutaneous tissue of the dorsal neck of each queen, then randomly removed after 7, 14, 28, 49, and 63 days after transplantation. Percentages of morphologically normal primordial and growing follicles (MNFs) were 88% and 97%, respectively, in fresh tissue samples (fresh controls), and 74% and 100%, respectively, immediately after thawing (cryo D0). No MNFs were found after 49 days of transplantation. In both fresh control and cryo D0 fragments, granulosa cells were frequently in proliferation. Two morphologically normal antral follicles were detected in one queen on Day 28 post-transplantation. Connective tissue fibers increased, suggesting replacement of active ovarian cortex by fibrous tissue. Tissue vascularization was observed at 7 days after grafting, and wide blood vessels were clearly visible on Days 49 and 63. In conclusion, although follicular survival was low after cryopreservation and grafting of cat ovarian tissue, follicles were able to develop up to the antral stage, which is an encouraging outcome.

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Section: Discussionsupporting

confidence: 92%

“…Although some attempts have been made to cryopreserve cat ovarian tissue [13][14][15][16][17][18][19][20][21], the protocols have not yet been well established. Working towards this goal, we have been studying techniques of cryopreservation and restoration of cat ovarian tissue function in our laboratory [21][22][23].…”

Section: Introductionmentioning

confidence: 99%

Function of Cryopreserved Cat Ovarian Tissue after Autotransplantation

Vilela

Leonel

Gonçalves

et al. 2019

Animals

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“…Trehalose and sucrose are the most commonly used disaccharides in cell and tissue cryopreservation studies (Baert et al 2013;Mouttham and Comizzoli 2016;Yokonishi and Ogawa 2016). Trehalose is the most efficient non-PCPA in inhibiting ice crystal formation during cryopreservation (Lee et al 2014a;Sambu 2015).…”

Section: Discussionmentioning

confidence: 99%

Effect of sucrose on cryopreservation of pig spermatogonial stem cells

Pan

Zhang

et al. 2017

Journal of Integrative Agriculture

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Sucrose is known to play an important role in the cryopreservation of sperm and female gonads; however, its effect on the cryopreservation of pig spermatogonial stem cells (pSSCs) has not been tested. The aim of this work was to study the effect of sucrose during pSSC cryopreservation and to find the most effective concentration in freezing medium. pSSCs were cryopreserved with freezing media containing different concentrations of sucrose (70, 140, 210, and 280 mmol L-1) and a control group without sucrose. The survival rates, plasma membrane integrity, and mitochondrial membrane potential of thawed cells were detected by trypan blue (TB) staining, SYBR-14/propidium iodide (PI) dual staining, and JC-1 staining, respectively. All the staining results showed an obvious increase in cell survival in the sucrose-treated groups as compared to that in the control group, with the exception of 280 mmol L-1 sucrose. Moreover, the 210 mmol L-1 sucrose group yielded the highest survival rate among all the groups (P<0.05). The results of SYBR-14/PI dual staining and JC-1 staining were consistent with those of TB staining as above described. Quantitative real-time PCR (qRT-PCR) indicated that the mRNA levels of three apoptosis-promoting genes (BAX, APAF1 and CASPASE9) were significantly higher in thawed cells than in cells before freezing (P<0.05). Moreover, the mRNA level of one anti-apoptotic gene (XIAP) was significantly lower in thawed cells than in cells before freezing (P<0.05). When comparing the mRNA expression of apoptosis-related genes in thawed cells, the mRNA level of the anti-apoptotic genes in the control group was significantly lower than that in the sucrose-treated groups (P<0.05). Western blot analyses showed that the expression levels of cleaved CASPASE9, CASPASE3 and PARP-1 in the sucrose-treated groups were lower than those in the control group and were the lowest in the 210 mmol L-1 sucrose group. Both qRT-PCR and Western blot analyses suggested that sucrose inhibited cell apoptosis during freezing and thawing. Briefly, sucrose promoted pSSCs survival after freezing and thawing, especially at a concentration of 210 mmol L-1 , which possibly assisted pSSC dehydration and inhibited cell apoptosis. These findings hold great promise for further studies of the regulatory mechanism of proliferation and differentiation of pSSCs.

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“…It has been reported that exposure of follicles to cryoprotectant agents could compromise their rates of survival (Rodrigues et al., ,b), even without freezing (Tsuribe, Gobbo, & da Cruz Landim‐Alvarenga, ). To improve vitrification, some modifications have been applied by researchers, which were the changes in the concentration and type of cryoprotectant (Herraiz et al., ; Lunardi et al., ; Yamaji et al., ), time (Ting, Yeoman, Lawson, & Zelinski, ), temperature (Mouttham & Comizzoli, ), the device used (Herraiz et al., ; Lunardi et al., ) and tissue size (Lu et al., ). Here, we noticed that follicular viability as a whole has been reduced in all of the groups after two weeks of tissue culture.…”

Section: Discussionmentioning

confidence: 99%

Comparison of the effects of Ham'sF10 and αMEM in combination with FBS or BSA in vitrification/warming solution on quality and viability of sheep ovarian follicles

Mohammadzadeh

Safdarian

Amidi

et al. 2017

Reprod Domestic Animals

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The aim of this study was to evaluate the effects of the two types of media, namely minimum essential medium (αMEM) and Ham'sF10, supplemented with foetal bovine serum (FBS) or bovine serum albumin (BSA) in vitrification/warming solution on the quality and viability of sheep ovarian follicles. Vitrification method was applied for cryopreservation of sheep ovarian cortex using Ham'sF10 and αMEM supplemented with either BSA or FBS. There were five groups: Fresh, Ham'sF10+ BSA, Ham'sF10+ FBS, αMEM + BSA and αMEM + FBS. Samples were cultured for two weeks after warming. Viability and morphology of follicles and DNA fragmentation in follicles and in tissue stroma cells were analysed before vitrification/warming and following one and two weeks of culture. The Ham'sF10+ FBS and Ham'sF10+ BSA groups showed a significant decrease in follicular viability after one week of culture (p < .05 vs. Fresh). Following two weeks of culture, all groups revealed a considerable fall in the number of viable follicles (p < .05 vs. Fresh). There was an increase in DNA fragmentation of connective tissue cells but not in the follicles (p < .05). Our results showed the better application of αMEM supplemented with BSA as a vitrification solution in improvement of cryopreservation effects and maintenance of follicular survival.

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The preservation of vital functions in cat ovarian tissues during vitrification depends more on the temperature of the cryoprotectant exposure than on the sucrose supplementation

Cited by 34 publications

References 53 publications

Function of Cryopreserved Cat Ovarian Tissue after Autotransplantation

Function of Cryopreserved Cat Ovarian Tissue after Autotransplantation

Effect of sucrose on cryopreservation of pig spermatogonial stem cells

Comparison of the effects of Ham'sF10 and αMEM in combination with FBS or BSA in vitrification/warming solution on quality and viability of sheep ovarian follicles

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