Ultrastructural markers of quality in human mature oocytes vitrified using cryoleaf and cryoloop

Sa, Nottola; Coticchio, Giovanni; Sciajno, Raffaella; Gambardella, A; Maione, M.; Scaravelli, Giulia; Bianchi, Serena; Macchiarelli, Guido; Borini, Andrea

doi:10.1016/s1472-6483(10)60280-5

Cited by 83 publications

(104 citation statements)

References 60 publications

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“…There were 44, 41 and 42 oocytes survived in 0 h, 2 h and 4 h group, respectively. According to Nottola SA et al's report [14], good vitrification protocol shouldn't affect the oocytes shape, ooplasm consistency or vacuole formation. Only survival oocytes which have intact zona pellucida (ZP) and plasmalemma, no overflow or shrinkage of cytoplasm, and no obviously cytoplasm vacuoles (diameter>5 μm, approximately) were analyzed after stained.…”

Section: Resultsmentioning

confidence: 99%

Effect of vitrification on mitochondrial membrane potential in human metaphase II oocytes

Han¹,

Liu²,

Wang

et al. 2012

J Assist Reprod Genet

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Section: Resultsmentioning

confidence: 99%

Effect of vitrification on mitochondrial membrane potential in human metaphase II oocytes

Han¹,

Liu²,

Wang

et al. 2012

J Assist Reprod Genet

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“…Nottola et al [24] also documented that 30% of oocytes (vitrified, human M-IIs) displayed a nonuniform distribution of microvilli at the oolemma. This abnormality was discussed in relation to possible disturbances in the sub-oolemma cytoskeletal network, and with the microfilament-richness of microvilli [24]. Patchy regions that were poor in microvilli were reported in human M-II oocytes that were slow-frozen with ethylene glycol and, interestingly, microvilli distribution at the oolemma was uniform with propanediol [37].…”

Section: Discussionmentioning

confidence: 95%

“…In a previous study analyzing various slow-freezing protocols in M-II human oocytes, an interrupted cortical actin network was reported in 11-26% of oocytes [36]. Nottola et al [24] also documented that 30% of oocytes (vitrified, human M-IIs) displayed a nonuniform distribution of microvilli at the oolemma. This abnormality was discussed in relation to possible disturbances in the sub-oolemma cytoskeletal network, and with the microfilament-richness of microvilli [24].…”

Section: Discussionmentioning

confidence: 95%

“…Past clinical studies with GV oocytes from a variety of sources as well as freezing protocols have documented wide ranges for survival, maturation, fertilization, and pre-implantation development (slow-freezing: [7][8][9][10][11][12]; vitrification: [13][14][15][16]). For mature oocytes, advancements in protocols have resulted in encouraging outcomes, including slow-freezing with choline-substitution [17][18][19][20] and vitrification with the Cryoleaf system [15,[21][22][23][24]. However, there are limited prior reports of either cholinebased slow-freezing [11,25] or vitrification with the Cryoleaf [15] in human GV oocytes, with no previous assessments of spindle integrity or chromosome alignment.…”

Section: Introductionmentioning

confidence: 99%

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Maturation outcomes are improved following Cryoleaf vitrification of immature human oocytes when compared to choline-based slow-freezing

Combelles

Ceyhan

Wang

et al. 2011

J Assist Reprod Genet

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Purpose The cryopreservation of immature oocytes permits oocyte banking for patients at risk of losing their fertility. However, the optimum protocol for such fertility preservation remains uncertain. Methods The present study investigated the survival, maturation, cytoskeletal and chromosome organization of sibling immature oocytes leftover from controlled ovarian stimulation cycles, that were either slow-frozen (with choline-substitution) or vitrified. A comparison group included oocytes that were never cryopreserved. Results Among the three groups, comparable rates were observed for both survival (67-70%) and polar body extrusion (59-79%). Significantly more oocytes underwent spontaneous activation after IVM following slow-freezing compared with either vitrification or no cryopreservation. Likewise, the incidence of spindle abnormalities was greatest in the slow-frozen group, with no differences in spindle morphometrics or chromosome organization.Conclusions While the overall incidence of mature oocytes with normal bipolar spindles from warmed immature oocytes was low, the yield using Cryoleaf vitrification was slightly superior to choline-based slow-freezing.

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“…Cytoplasmic vacuolization is one of the most common signs of damage in cryopreserved oocytes (Nottola et al 2009;Coticchio et al 2010) and ovarian tissue ) and has been adopted as a specific marker for cryodamage (Coticchio et al 2010). Silva et al (2000) have suggested that damage to smooth endoplasmic reticulum (ER) leads to vacuolization.…”

Section: Introductionmentioning

confidence: 99%

Vitamin E-analog Trolox prevents endoplasmic reticulum stress in frozen-thawed ovarian tissue of capuchin monkey (Sapajus apella)

Brito

Scalercio

et al. 2013

Cell Tissue Res

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Ovarian fragments were exposed to 0.5 M sucrose and 1 M ethylene glycol (freezing solution; FS) with or without selenium or Trolox. Histological and ultrastructural analyses showed that the percentages of normal follicles in control tissue and in tissue after exposure to FS+50 μM Trolox were similar. Trolox prevented endoplasmic reticulum (ER)-related vacuolization, which is commonly observed in oocytes and stromal tissue after exposure to FS. From the evaluated stress markers, superoxide dismutase 1 (SOD1) was up-regulated in ovarian tissue exposed to FS+10 ng/ml selenium. Ovarian fragments were subsequently frozenthawed in the presence of FS with or without 50 μM Trolox, followed by in vitro culture (IVC). Antioxidant capacity in ovarian fragments decreased after freeze-thawing in Troloxfree FS compared with FS+50 μM Trolox. Although freezing itself minimized the percentage of viable follicles in each solution, Trolox supplementation resulted in higher rates of viable follicles (67 %), even after IVC (61 %). Furthermore, stress markers SOD1 and ERp29 were up-regulated in ovarian tissue frozen-thawed in Trolox-free medium. Relative mRNA expression of growth factors markers was evaluated after freeze-thawing followed by IVC. BMP4, BMP5, CTGF, GDF9 and KL were down-regulated independently of the presence of Trolox in FS but down-regulation was less pronounced in the presence of Trolox. Thus, medium supplementation with 50 μM Trolox prevents ER stress and, consequently, protects ovarian tissue from ER-derived cytoplasmic vacuolization. ERp29 but not ERp60, appears to be a key marker linking stress caused by freezing-thawing and cell vacuolization.

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Ultrastructural markers of quality in human mature oocytes vitrified using cryoleaf and cryoloop

Cited by 83 publications

References 60 publications

Effect of vitrification on mitochondrial membrane potential in human metaphase II oocytes

Effect of vitrification on mitochondrial membrane potential in human metaphase II oocytes

Maturation outcomes are improved following Cryoleaf vitrification of immature human oocytes when compared to choline-based slow-freezing

Vitamin E-analog Trolox prevents endoplasmic reticulum stress in frozen-thawed ovarian tissue of capuchin monkey (Sapajus apella)

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