Spontaneous in vitro maturation and artificial activation of human germinal vesicle oocytes recovered from stimulated cycles

Escrich, L.; Grau, N.; Mercader, Amparo; Rubio, Carmen; Pellicer, António; Escribá, Marı́a-José

doi:10.1007/s10815-010-9493-5

Cited by 26 publications

(25 citation statements)

References 41 publications

(81 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…All previous investigations support the findings of this study; specifically, LH stimulation, not FSH, contributes to the nuclear maturation of human GV-stage oocytes (17)(18)(19)(20)(21)(22)(23)(24). In terms of maturation time, GV oocytes progressed to MII during the first 23.70 AE 0.94 hours of IVM; this is in agreement with previous reports (25). Supplementation with FSH/LH to the IVM medium shortened the maturation time by approximately 1 hour (22.78 AE 0.87 h), although this difference was not statistically significant.…”

Section: Figuresupporting

confidence: 93%

In vitro maturation of cumulus–partially enclosed immature human oocytes by priming with gonadotropin

Yan

et al. 2011

Fertility and Sterility

View full text Add to dashboard Cite

Section: Figuresupporting

confidence: 93%

In vitro maturation of cumulus–partially enclosed immature human oocytes by priming with gonadotropin

Yan

et al. 2011

Fertility and Sterility

View full text Add to dashboard Cite

“…Higher maturation rates can be achieved by supplementing the maturation media with gonadotropins (80 %, [9]). The inadequacy of the in vitro system we employed is also evident in the low percentage of IVM oocytes that displayed a normal activation response following parthenogenetic activation (41.0 %), especially in comparison to results obtained with in vivo matured oocytes (76 %; [6]). …”

Section: Discussionmentioning

confidence: 91%

“…Large cumulus-oocyte-complexes were collected (Fig. 1a), washed and individually cultured in 50 μL CCM (Vitrolife, Göteborg, Sweden) under standard conditions (37°C, 5%CO 2 in air) for 48 h, as in the case of GV oocytes recovered from stimulated cycles [6,7].…”

Section: Methodsmentioning

confidence: 99%

“…MII oocytes ( Fig. 1c and d) that were healthy in appearance were artificially activated with calcium ionophore (A23184) and puromycin [6,14]. In short, oocytes were exposed to A23187 (8 mM, Sigma, Barcelona, Spain) for 5 min and subsequently cultured in puromycin (10 μg/mL; Sigma, Barcelona, Spain) for 5 h. Once activated, eggs were cultured in 50 μL IVF medium (Vitrolife, Göteborg, Sweden) in the previously referred conditions for 16-20 h and then assessed for extrusion of the second polar body (2PB) and number of pronuclei.…”

Section: Methodsmentioning

confidence: 99%

See 1 more Smart Citation

Spontaneous in vitro maturation of oocytes prior to ovarian tissue cryopreservation in natural cycles of oncologic patients

Escribá

Grau

Escrich

et al. 2012

J Assist Reprod Genet

Self Cite

View full text Add to dashboard Cite

Purpose To evaluate the recovery rate and spontaneous in vitro maturation (IVM) of immature oocytes enclosed within or released from follicles during the processing of ovarian tissue prior to its cryopreservation. Methods Thirty-three oncologic patients who had not previously undergone chemo or radiotherapy underwent ovarian tissue cryopreservation (OTC) during natural menstrual cycles. Immature oocytes, enclosed within follicles or released during ovarian cortex processing, were collected and matured spontaneously in vitro for 48 h. Nuclear maturation was assessed every 24 h and the ability of the IVM oocytes to display a normal activation response following parthenogenetic activation was evaluated. The following outcome measures were also evaluated: disease, age, FSH, LH, E2, P4 and AMH serum levels, menstrual cycle day, recovery and spontaneous IVM and parthenogenetic activation rates. Results Oocytes recovered per patient were 3.3±0.7 (1.8-4.7 oocytes, 95CI), regardless of the menstrual phase. The mean number of IVM oocytes per patient was 1.3±0.2 oocytes (95CI: 0.8-1.8), regardless of menstrual phase (p0 0.86) and oocyte origin (p00.61). Forty-one percent of oocytes extruded the second polar body and formed one pronucleus after parthenogenetic activation. Conclusion Twenty-one of the 33 women (63.6 %) requesting OTC produced at least one mature oocyte.

show abstract

“…To avoid the ethical issues involved in creating fertilized embryos for research purposes and the additional manipulation required to remove the pronuclei, we based our protocol on an established method for artificial activation of oocytes arrested in metaphase of the second meiotic division, or metaphase II (MII), with calcium ionophore (A23187) [9][10][11][12][13][14] . The activated MII oocytes resume meiosis and extrude the PB2.…”

Section: Development Of the Protocolmentioning

confidence: 99%

Generation of meiomaps of genome-wide recombination and chromosome segregation in human oocytes

et al. 2016

View full text Add to dashboard Cite

We have developed a protocol for the generation of genome-wide maps (meiomaps) of recombination and chromosome segregation for the three products of human female meiosis: the first and second polar bodies (PB1 and PB2) and the corresponding oocyte. PB1 is biopsied and the oocyte is artificially activated by exposure to calcium ionophore, after which PB2 is biopsied and collected with the corresponding oocyte. The whole genomes of the polar bodies and oocytes are amplified by multiple displacement amplification and, together with maternal genomic DNA, genotyped for ∼300,000 single-nucleotide polymorphisms (SNPs) genome-wide by microarray. Informative maternal heterozygous SNPs are phased using a haploid PB2 or oocyte as a reference. A simple algorithm is then used to identify the maternal haplotypes for each chromosome, in all of the products of meiosis for each oocyte. This allows mapping of crossovers and analysis of chromosome segregation patterns. The protocol takes a minimum of 3-5 d and requires a clinical embryologist with micromanipulation experience and a molecular biologist with basic bioinformatic skills. It has several advantages over previous methods; importantly, the use of artificial oocyte activation avoids the creation of embryos for research purposes. In addition, compared with next-generation sequencing, targeted SNP genotyping is cost-effective and it simplifies the bioinformatic analysis, as only one haploid reference sample is required to establish phase for maternal haplotyping. Finally, meiomapping is more informative than copy-number analysis alone for analysis of chromosome segregation patterns. Using this protocol, we have provided new insights that may lead to improvements in assisted reproduction for the treatment of infertility.

show abstract

Spontaneous in vitro maturation and artificial activation of human germinal vesicle oocytes recovered from stimulated cycles

Abstract: GV spontaneously mature at the nuclear level. Their NA are haploid, but their cytoplasmic competence is compromised. Variables were not found to be predictors of oocyte competence, probably due to our population being homogeneous with respect to most of the variables studied.

Cited by 26 publications

References 41 publications

In vitro maturation of cumulus–partially enclosed immature human oocytes by priming with gonadotropin

In vitro maturation of cumulus–partially enclosed immature human oocytes by priming with gonadotropin

Spontaneous in vitro maturation of oocytes prior to ovarian tissue cryopreservation in natural cycles of oncologic patients

Generation of meiomaps of genome-wide recombination and chromosome segregation in human oocytes

Contact Info

Product

Resources

About