Cryopreservation of Mouse Ovarian Tissue Following Prolonged Exposure to an Ischemic Environment

Cleary, Michelle Lee; Snow, Melanie Jennifer; Paris, M.C.J.; Shaw, Jillian M; Cox, Shae-Lee; Jenkin, Graham

doi:10.1006/cryo.2001.2315

Cited by 36 publications

(17 citation statements)

References 41 publications

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“…The vials were plunged directly into liquid nitrogen (−196 °C) and stored for up to 5 days before thawing. The freezing curve used in this study was chosen because most procedures currently used to cryopreserve oocytes [25], [26], [27], [28], [29] and [30] and ovarian tissue [10], [11], [12], [14], [15], [18], [24], [31], [32] and [33] stipulate a cooling rate of 0.3-0.5 °C/min from the seeding temperature (usually −5 to −9 °C) to a lower temperature, usually between −30 and −40 °C.…”

Section: Freezing and Thawing Proceduresmentioning

confidence: 99%

Cryopreservation of caprine ovarian tissue using glycerol and ethylene glycol

et al. 2004

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Cryopreservation of ovarian tissue may be a potential alternative for the conservation of genetically superior animals, including high milk-and meat-producing goat breeds. However, until now, no information was available concerning the cryopreservation of preantral follicles (PF) enclosed in caprine ovarian tissue. The objective of the present study was to evaluate the structural and ultrastructural characteristics of caprine PF after exposure to and cryopreservation of ovarian tissue in 1.5 and 3 M glycerol (GLY) and ethylene glycol (EG). At the slaughterhouse, each ovarian pair from five adult mixed breed goats was divided into nine fragments and randomly distributed into treatment groups. One fragment was immediately fixed for histological examination and ultrastructural analysis, after slaughter (control). Four of the ovarian fragments were equilibrated at 20 °C for 20 min in 1.8 ml of MEM containing 1.5 or 3 M GLY or EG for a toxicity test and the final four fragments were slowly frozen using these cryoprotectants at the concentrations above. After toxicity testing and freezing/thawing, the ovarian fragments were fixed for histological examination. Histological analysis showed that after toxicity testing and cryopreservation of the ovarian tissue in GLY or EG at both concentrations, the percentage of normal PF was significantly lower than controls. Ultrastructural analysis of PF frozen in 1.5 and 3 M GLY, as well as 3 M EG demonstrated that these follicles remained morphologically normal. In conclusion, we demonstrated cryopreservation of caprine PF in ovarian tissue.

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Section: Freezing and Thawing Proceduresmentioning

confidence: 99%

Cryopreservation of caprine ovarian tissue using glycerol and ethylene glycol

et al. 2004

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“…Nevertheless, Gook et al also observed a better follicular preservation when using a slow freezing rate (0.3°C/min) with human ovarian tissue (Gook et al, 1999). Whereas Cleary et al observed no difference in terms of follicular growth after grafting, between a conventional embryo freezing protocol (0.3°C/min) and a passive cooling at 1°C/min from 0°C to -84°C on the mouse ovarian tissue (Cleary et al, 2001). Although these two cooling rates (0.3°C/min and 2°C/min) could be considered as slow, these results may be explained by a difference in cell dehydration during the post-seeding step.…”

Section: Discussionmentioning

confidence: 96%

New Approaches of Ovarian Tissue Cryopreservation from Domestic Animal Species

Neto¹,

Joly²,

Commin³

et al. 2012

Current Frontiers in Cryopreservation

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“…Cryopreservation of the ovarian tissue with slow freezing method and thawing Cryopreservation was performed according to previous reports [10,16,18]. Ovarian cortical fragments were transferred from holding medium to cryopreservation medium contained with L-15 medium supplemented with 1.5 M DMSO (Fluka Chemie GmbH, Buchs, Spain) and 0.1 M sucrose for 5 min at 4°C.…”

Section: Ovarian Tissue Culturementioning

confidence: 99%

“…In association with the freezing techniques, culture of the ovarian tissue in vitro is a fundamental approach to reanimate the tissue and grow more oocytes to an advanced stage [5,7,8]. However, there may be several hours between the ovarian tissue collection and the cryopreservation process itself which compromises (through ischemia and oxidative stress) the viability and developmental competence of the germ cells within the gonads [3,9,10]. During this critical period, cellular respiration in mitochondria is still occurring despite the ischemic environment [10][11][12].…”

Section: Introductionmentioning

confidence: 99%

“…However, there may be several hours between the ovarian tissue collection and the cryopreservation process itself which compromises (through ischemia and oxidative stress) the viability and developmental competence of the germ cells within the gonads [3,9,10]. During this critical period, cellular respiration in mitochondria is still occurring despite the ischemic environment [10][11][12]. Thus, reversibly reducing the metabolic activity of the cells (by using a metabolic disrupting agent) is an interesting option for protecting and prolonging the tissue integrity.…”

Section: Introductionmentioning

confidence: 99%

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Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP) pre-exposure ensures follicle integrity during in vitro culture of ovarian tissue but not during cryopreservation in the domestic cat model

Tanpradit

Chatdarong

Comizzoli

2016

J Assist Reprod Genet

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Purpose Temporary and reversible downregulation of metabolism may improve the survival of tissues exposed to nonphysiological conditions during transport, in vitro culture, and cryopreservation. The objectives of the study were to (1) optimize the concentration and duration of carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP-a mitochondrial uncoupling agent) exposures for biopsies of domestic cat ovarian tissue and (2) examine the effects of FCCP preexposures on follicle integrity after tissue culture and/or cryopreservation. Methods Biopsies of cat ovarian tissue were first treated with various concentrations of FCCP (0, 10, 40, or 200 nM) for 10 or 120 min to determine the most suitable pre-exposure conditions. Based on these results, tissues were pre-exposed to 200 nM FCCP for 120 min for the subsequent studies on culture and cryopreservation. In all experiments and for each treatment group, tissue activity and integrity were measured by mitochondrial membrane potential (relative optical density of rhodamine 123 fluorescence), follicular viability (calcein assay), follicular morphology (histology), granulosa cell proliferation (Ki-67 immunostaining), and follicular density.Results Ovarian tissues incubated with 200 nM FCCP for 120 min led to the lowest mitochondrial activity (1.17 ± 0.09; P < 0.05) compared to control group (0 nM; 1.30 ± 0.12) while maintaining a constant percentage of viable follicles (75.3 ± 7.8 %) similar to the control group (71.8 ± 11.7 %; P > 0.05). After 2 days of in vitro culture, percentage of viable follicles (78.8 ± 8.9 %) in similar pre-exposure conditions was higher (P < 0.05) than in the absence of FCCP (61.2 ± 12.0 %) with percentages of morphologically normal follicles (57.6 ± 17.3 %) not different from the fresh tissue (70.2 ± 7.1 %; P > 0.05). Interestingly, percentages of cellular proliferation and follicular density were unaltered by the FCCP exposures. Based on the indicators mentioned above, the FCCP-treated tissue fragments did not have a better follicle integrity after freezing and thawing. Conclusions Pre-exposure to 200 nM FCCP during 120 min protects and enhances the follicle integrity in cat ovarian tissue during short-term in vitro culture. However, FCCP does not appear to exert a beneficial or detrimental effect during ovarian tissue cryopreservation.

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Cryopreservation of Mouse Ovarian Tissue Following Prolonged Exposure to an Ischemic Environment

Cited by 36 publications

References 41 publications

Cryopreservation of caprine ovarian tissue using glycerol and ethylene glycol

Cryopreservation of caprine ovarian tissue using glycerol and ethylene glycol

New Approaches of Ovarian Tissue Cryopreservation from Domestic Animal Species

Carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP) pre-exposure ensures follicle integrity during in vitro culture of ovarian tissue but not during cryopreservation in the domestic cat model

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