Cryopreservation of Unfertilized Mouse Oocytes: The Effect of Replacing Sodium with Choline in the Freezing Medium

Stachecki, James J.; Cohen, Jacques; Willadsen, S. M.

doi:10.1006/cryo.1998.2130

Cited by 119 publications

(52 citation statements)

References 21 publications

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“…With the advent of improved protocols [4,34,35], novel ultrastructural evidence was generated. Comparing fresh oocytes with others frozen with a CRSC involving 0.1 mol/l sucrose as an extracellular CPA, Ghetler et al [36] found massive reduction in the number of CG as a effect of cryopreservation, concluding that stored oocytes should be microinjected rather than inseminated by standard IVF to prevent possible fertilization failure secondary to zona hardening.…”

Section: Discussionmentioning

confidence: 99%

Qualitative and morphometric analysis of the ultrastructure of human oocytes cryopreserved by two alternative slow cooling protocols

Coticchio¹,

Borini²,

Distratis³

et al. 2010

J Assist Reprod Genet

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Purpose To ascertain possible cell damage from cryopreservation, the ultrastructure of human oocytes cryopreserved by slow cooling was assessed. Materials and methods Cryopreservation was performed through two protocols with one-step or two-step propanediol. Fresh control oocytes were examined for comparison. Samples were processed for transmission electron microscopy analysis. Results By light microscopy, both fresh and frozen-thawed oocytes appeared regularly rounded, with intact zona pellucida, and homogeneous cytoplasm. By electron microscopy observation, organelles were abundant and uniformly dispersed. Mitochondria-smooth endoplasmic reticulum associations appeared regular. However, both the amount and density of cortical granules appeared abnormally reduced in frozen-thawed samples. Slight to moderate vacuolization was also found in the ooplasm of oocytes of both frozen groups. Conclusions Slow cooling ensures a good overall preservation of human oocytes. However, cytoplasmic vacuolization and cortical granule loss appears associated with cryopreservation, irrespective of the protocol used.

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Section: Discussionmentioning

confidence: 99%

Qualitative and morphometric analysis of the ultrastructure of human oocytes cryopreserved by two alternative slow cooling protocols

Coticchio¹,

Borini²,

Distratis³

et al. 2010

J Assist Reprod Genet

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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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“…Attempts to improve outcome by altering the components of the freezing extender (e.g. replacing sodium chloride with choline chloride; Stachecki et al, 1998a;Stachecki et al, 1998b;Quintans et al, 2002) suggest that there is still some room for improvements in the standard techniques widely in use. Vitrification usually exposes the oocytes to substantially higher concentration of cryoprotectants, in the range of 5.0 to 7.0 M, and cryopreservation is done at cooling rates of 2,500ºC per minute or more, depending on the technique used.…”

Section: Oocyte Cryopreservationmentioning

confidence: 99%

Genome Banking for Vertebrates Wildlife Conservation

Saragusty¹

2012

Current Frontiers in Cryobiology

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Cryopreservation of Unfertilized Mouse Oocytes: The Effect of Replacing Sodium with Choline in the Freezing Medium

Cited by 119 publications

References 21 publications

Qualitative and morphometric analysis of the ultrastructure of human oocytes cryopreserved by two alternative slow cooling protocols

Qualitative and morphometric analysis of the ultrastructure of human oocytes cryopreserved by two alternative slow cooling protocols

Cryopreservation of caprine ovarian tissue using glycerol and ethylene glycol

Genome Banking for Vertebrates Wildlife Conservation

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