Confocal microscopic analysis of the spindle and chromosome configurations of human oocytes matured in vitro

Li, Yuan; Feng, Hua; Cao, Yi-Juan; Zheng, Guangjuan; Yang, Yong; Mullen, Steven F.; Critser, John K.; Chen, Zi‐Jiang

doi:10.1016/j.fertnstert.2005.06.064

Cited by 145 publications

(98 citation statements)

References 37 publications

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“…The presence of disorganized microtubules at the meiotic spindle seen in the present study has also been described in women oocytes (Li et al, 2006). Damage to microtubule organization results in depolymerization of the meiotic spindles in mammalian oocytes, leading to an irreversible process that results in chromosomal abnormalities after fertilization, such as aneuploidy and polyploidy (Wang and Sun, 2006).…”

Section: Discussionsupporting

confidence: 80%

Influence of equine growth hormone, insulin-like growth factor-I and its interaction with gonadotropins on in vitro maturation and cytoskeleton morphology in equine oocytes

Pereira

Lorenzo

Carneiro

et al. 2013

Animal

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In horses, successful in vitro fertilization procedures are limited by our inability to consistently mature equine oocytes by in vitro methods. Growth hormone (GH) is an important regulator of female reproduction in mammals, playing an important role in ovarian function, follicular growth and steroidogenesis. The objectives of this research were to investigate: the effects of equine growth hormone (eGH) and insulin-like growth factor-I (IGF-I) on the in vitro maturation (IVM) of equine oocytes, and the effects of eGH in addition to estradiol (E 2 ), gonadotropins (FSH and LH) and fetal calf serum (FCS) on IVM. We also evaluated the cytoskeleton organization of equine oocytes after IVM with eGH. Equine oocytes were aspirated from follicles ,30 mm in diameter and matured for 30 h at 38.58C in air with 5% CO 2 . In experiment 1, selected cumulus-oocyte complexes (COCs) were randomly allocated as follows: (a) control (no additives); (b) 400 ng/ml eGH; (c) 200 ng/ml IGF-I; (d) eGH 1 IGF-I; and (e) eGH 1 IGF-I 1 200 ng/ml anti-IGF-I. In addition to these treatment groups, we also added 1 mg/ml E 2 , 5 IU/ml FSH, 10 IU/ml LH and 10% FCS in vitro (experiment 2). Oocytes were stained with markers for microtubules (anti-a-tubulin antibody), microfilaments (AlexaFluor 488 Phalloidin) and chromatin (TO-PRO 3 -iodide) and assessed via confocal microscopy. No difference was observed when eGH and IGF-I was added into our IVM system. However, following incubation with eGH alone (40%) and eGH, E 2 , gonadotropins and FCS (36.6%) oocytes were classified as mature v. 17.6% of oocytes in the control group (P , 0.05). Matured equine oocytes showed that a thin network of filaments concentrated within the oocyte cortex and microtubules at the metaphase spindle showed a symmetrical barrel-shaped structure, with chromosomes aligned along its midline. We conclude that the use of E 2 , gonadotropins and FCS in the presence of eGH increases the number of oocytes reaching oocyte competence.

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Section: Discussionsupporting

confidence: 80%

Influence of equine growth hormone, insulin-like growth factor-I and its interaction with gonadotropins on in vitro maturation and cytoskeleton morphology in equine oocytes

Pereira

Lorenzo

Carneiro

et al. 2013

Animal

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“…One concern with the approach is the relatively high rate of developmental incompetence observed in oocytes subject to IVM. Li et al (2006) raised concerns regarding possible deleterious effects that IVM might have on the organisation of the meiotic spindle and chromosomal alignment. Although, at present, there are no indications of increased risk of congenital malformations in children conceived by IVM, the processes involved and the longterm outcomes are still poorly understood (Reinblatt & Buckett 2008), and data from ongoing follow-up studies are awaited.…”

mentioning

confidence: 99%

The impact of ovarian stimulation for IVF on the developing embryo

Santos¹,

Kuijk²,

Macklon³

2010

Reproduction

171

126

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The use of assisted reproductive technologies (ART) has been increasing over the past three decades, and, in developed countries, ART account for 1-3% of annual births. In an attempt to compensate for inefficiencies in IVF procedures, patients undergo ovarian stimulation using high doses of exogenous gonadotrophins to allow retrieval of multiple oocytes in a single cycle. Although ovarian stimulation has an important role in ART, it may also have detrimental effects on oogenesis, embryo quality, endometrial receptivity and perinatal outcomes. In this review, we consider the evidence for these effects and address possible underlying mechanisms. We conclude that such mechanisms are still poorly understood, and further knowledge is needed in order to increase the safety of ovarian stimulation and to reduce potential effects on embryo development and implantation, which will ultimately be translated into increased pregnancy rates and healthy offspring.

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“…It was suggested that the presence of a birefringent spindle in MII oocytes could predict a higher fertilization rate and embryo developmental competence after ICSI (25). It is also well recognized that the spindle and chromosomes have critical importance in fertilization and subsequent embryo development (26). Therefore, spindle imaging could help to identify abnormalities in the oocyte MS (27).…”

Section: Discussionmentioning

confidence: 99%

Analysis of meiotic spindle and zona pellucida birefringenceof IVM oocytes in PCOS patients

et al. 2017

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Background/aim: Polycystic ovarian syndrome (PCOS) is one of the most common causes of infertility. One of the best therapeutic options for PCOS patients is intracytoplasmic sperm injection (ICSI). In vitro maturation (IVM) can also be a useful technique for these women. The goal of this study was to evaluate both the zona pellucida (ZP) birefringence and meiotic spindle (MS) of in vivo-and in vitro-matured oocytes from PCOS patients using the PolScope system.Materials and methods: Immature oocytes undergoing IVM and MII oocytes were obtained from PCOS patients in an ICSI program. Using PolScope, the presence of MS and ZP birefringence was assessed in both in vivo-matured oocytes (n = 32) and IVM oocytes (n = 24). Oocytes were classified as having highly birefringent (HB) ZP and lowly birefringent (LB) ZP. Furthermore, the rates of fertilization after ICSI were evaluated. Results:The maturation rate was 68.5% after IVM. The percentage of a HB-ZP was significantly higher in the IVM oocytes than in vivomatured ones (58.3% vs. 31.2%, respectively; P = 0.04). There were similar outcomes for the fertilization rates and MS detection between the two groups (P = 0.80 and P = 0.53, respectively). Conclusion:Clinical IVM is a safe technology for the maturation and maintenance of oocyte integrity in PCOS patients. The use of the noninvasive PolScope is recommended for detection of healthy oocytes in ICSI.

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Confocal microscopic analysis of the spindle and chromosome configurations of human oocytes matured in vitro

Cited by 145 publications

References 37 publications

Influence of equine growth hormone, insulin-like growth factor-I and its interaction with gonadotropins on in vitro maturation and cytoskeleton morphology in equine oocytes

Influence of equine growth hormone, insulin-like growth factor-I and its interaction with gonadotropins on in vitro maturation and cytoskeleton morphology in equine oocytes

The impact of ovarian stimulation for IVF on the developing embryo

Analysis of meiotic spindle and zona pellucida birefringenceof IVM oocytes in PCOS patients

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