The effects of ultrarapid freezing on meiotic and mitotic spindles of mouse oocytes and embryos

Sathananthan, A. Henry; Ng, Soon-Chye; Trounson, Alan; Bongso, Ariff; Ratnam, S. S.; Ho, Jean K.; Mok, Helen; Lee, M. N.

doi:10.1002/mrd.1120210407

Cited by 97 publications

(35 citation statements)

References 15 publications

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“…In addition, there was no difference in the types of damage observed within various groups. Similar types of damage have been observed in earlier studies (Sathananthan et al, 1988;Dhali et al, 2000;Albarracin et al, 2005a,b). Post-thaw survivability of buffalo oocytes in this study was 37.6-61.3%, with survivability increasing with time in IVM prior to vitrification, but survivability was lower in all vitrified groups than that of non-vitrified oocytes (95.5%).…”

Section: Discussionsupporting

confidence: 75%

Effect of Meiotic Stages During In Vitro Maturation on the Survival of Vitrified-Warmed Buffalo Oocytes

Sharma

Loganathasamy

2007

Vet Res Commun

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The present study was conducted to investigate the effect of meiotic stages during in vitro maturation (IVM) on the survival of vitrified-warmed buffalo oocytes, vitrified at different stages of IVM. Cumulus oocyte complexes obtained from slaughterhouse ovaries were randomly divided into 6 groups: control (non-vitrified, matured for 24 h at 38 +/- 1 degrees C, 5% CO2 in humidified air), and those matured for 0 h (vitrified before IVM) or 6, 12, 18 and 24 h before vitrification. Cumulus oocyte complexes were vitrified in solution consisting of 40% w/v propylene glycol and 0.25 mol/L trehalose in phosphate-buffered saline supplemented with 4% w/v bovine serum albumin. Vitrified cumulus oocyte complexes were stored at -196 degrees C (liquid nitrogen) for at least 7 days and then thawed at 37 degrees C; cryoprotectant was removed with 1 mol/L sucrose solution. Cumulus oocyte complexes in the 0, 6, 12, 18 and 24 h groups were then matured for an additional 24, 18, 12, 6 and 0 h, respectively, to complete 24 h of IVM. Among the five vitrification groups, 89-92% of cumulus oocyte complexes were recovered, after warming, of which 84-91% were morphologically normal. Overall survivability of vitrified cumulus oocyte complexes was lower (p < 0.05) than that of non-vitrified cumulus oocyte complexes (94.5%). Survival rates of cumulus oocyte complexes matured 24 h prior to vitrification (61.3%) were higher (p < 0.05) than those matured for 12 h (46.7%), 6 h (40.6%) and 0 h (37.6%). Nuclear status following 24 h IVM was assessed. A higher proportion of non-vitrified (control) oocytes (72.7%) reached metaphase II (M-II) stage in control than oocytes vitrified for 24 h (60.0%), 18 h (54.4), 12 h (42.3%), 6 h (33.3%) and 0 h (31.6%) (p < 0.05). The results suggest that length of time in maturation medium prior to vitrification influences post-thaw survivability of buffalo oocytes; longer intervals resulted in higher survival rates.

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Section: Discussionsupporting

confidence: 75%

Effect of Meiotic Stages During In Vitro Maturation on the Survival of Vitrified-Warmed Buffalo Oocytes

Sharma

Loganathasamy

2007

Vet Res Commun

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“…In this respect, oocytes are susceptible to damage during cooling and/or freezing and thawing (Leibo 1980) because they are structurally complex and relatively impermeable to both water and cryoprotectants (CPAs; Agca et al 1998). While the specific molecular elements or pathways disrupted during oocyte cryopreservation are not well characterised, many ultrastructural elements critical to the maintenance of developmental competence are damaged during freezing and thawing (Parks & Ruffing 1992), including the plasma membrane (Ashwood-Smith et al 1988), the actin cytoskeleton and the meiotic spindle (Sathananthan et al 1988, Park et al 1997. Oocyte cryopreservation can also lead to 'zona hardening', as a result of CPA-induced premature cortical granule release (Ghetler et al 2006), and it is for this reason that cryopreserved human oocytes are usually fertilised by intracytoplasmic sperm injection rather than by conventional IVF (for reviews see Coticchio et al 2007, Gook & Edgar 2007.…”

Section: Introductionmentioning

confidence: 99%

Protective effects of the cumulus-corona radiata complex during vitrification of horse oocytes

Tharasanit

Colleoni²,

Galli³

et al. 2009

Reproduction

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Vitrifying oocytes is a potentially valuable means of preserving the female germ line, but significantly compromises oocyte developmental competence. This study examined the hypothesis that the cumulus complex protects the oocyte during vitrification. Vitrified-warmed immature cumulus oocyte complexes (COCs) were labelled with a plasma membrane impermeant DNA marker (ethidium homodimer-1) to examine the percentage and location of dead cumulus cells, and to investigate the effect of the proportion of dead cells (C1,C2 or C3) on the success of in vitro maturation (IVM). Further, oocytes were labelled for connexin-43 or injected with Lucifer yellow dye to determine whether the integrity of the gap junctions between an oocyte and its cumulus was compromised by vitrification. Finally, the effect of denuding immature and mature oocytes on their ability to withstand vitrification was examined. Cryopreserving immature COCs increased the number of dead cumulus cells (13 vs 2.6% for controls; P!0.05). However, an increased proportion of dead cumulus cells did not affect post-warming maturation rates (w30% MII ) presumably because dead cells were located at the periphery of the cumulus mass and cumulus-oocyte gap junction communication was not disrupted. Moreover, cumulus removal prior to IVM or vitrification indicated that while the cumulus does protect immature oocytes during vitrification it does so by mechanisms other than support during maturation. Cumulus presence was also found to protect mature equine oocytes against vitrification-induced damage since cumulus-enclosed MII oocytes preserved their meiotic spindle quality better during vitrification than denuded oocytes (38.1 vs 3.1% normal spindles; P!0.05).

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“…However, while live offspring have been produced using cryopreserved oocytes in a number of species (e.g. man: Chen 1986; mouse: Sathananthan et al 1988;cattle: Lim et al 1991;horse: Maclellan et al 2002), the overall success in terms of developmental competence is disappointing (Porcu 2001). It appears that the cryopreservability of oocytes is complicated by their structural complexity, low surface area:volume ratio and relatively low plasma membrane permeability to water and/or cryoprotectants (Leibo 1980, Agca et al 1998.…”

Section: Introductionmentioning

confidence: 99%

Effect of cumulus morphology and maturation stage on the cryopreservability of equine oocytes

Tharasanit¹,

Colleoni²,

Lazzari³

et al. 2006

Reproduction

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Oocyte cryopreservation is a potentially valuable way of preserving the female germ line. However, the developmental competence of cryopreserved oocytes is presently poor. This study investigated whether the morphology of the cumulus complex surrounding an immature equine oocyte and/or the oocyte's stage of maturation affect its cryopreservability. Compact (Cp) and expanded (Ex) cumulus oocyte complexes (COCs) were vitrified either shortly after recovery (germinal vesicle stage, GV) or after maturation in vitro (IVM); cryoprotectant-treated and -untreated non-frozen oocytes served as controls. In Experiment I, oocytes matured in vitro and then vitrified, or vice versa, were examined for maturation stage and meiotic spindle quality. Cp and Ex COCs vitrified at the GV stage matured at similar rates during subsequent IVM (41 vs 46% MII), but meiotic spindle quality was better for Cp than Ex (63 vs 33% normal spindles). Vitrifying oocytes after IVM resulted in disappointing post-warming spindle quality (32 vs 28% normal for Cp vs Ex). In Experiment II, oocytes from Cp and Ex COCs vitrified at the GV or MII stages were fertilized by intracytoplasmic sperm injection (ICSI) and monitored for cleavage and blastocyst formation. Oocytes vitrified prior to IVM yielded higher cleavage rates (34 and 27% for Cp and Ex COCs) than those vitrified after IVM (16 and 4%). However, only one blastocyst was produced from a sperm-injected vitrified-warmed oocyte (0.4 vs 9.3% and 13% blastocysts for cryoprotectant-exposed and -untreated controls). It is concluded that, when vitrification is the chosen method of cryopreservation, Cp equine COCs at the GV stage offer the best chance of an MII oocyte with a normal spindle and the potential for fertilization; however, developmental competence is still reduced dramatically. Reproduction (2006) 132 759-769

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The effects of ultrarapid freezing on meiotic and mitotic spindles of mouse oocytes and embryos

Cited by 97 publications

References 15 publications

Effect of Meiotic Stages During In Vitro Maturation on the Survival of Vitrified-Warmed Buffalo Oocytes

Effect of Meiotic Stages During In Vitro Maturation on the Survival of Vitrified-Warmed Buffalo Oocytes

Protective effects of the cumulus-corona radiata complex during vitrification of horse oocytes

Effect of cumulus morphology and maturation stage on the cryopreservability of equine oocytes

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