Optimal ICSI timing after the first polar body extrusion in in vitro matured human oocytes.

Background: This study was conducted to investigate the fertilization and embryo development of human oocytes injected at different time intervals after extrusion of the first polar body (PB) following in vitro maturation (IVM) in IVM cycles. Also, we evaluated whether spindle imaging could serve as a tool to determine the optimal ICSI time.

Methods: Oocytes were collected from 43 women with polycystic ovary syndrome.

New alternative to infertility treatment for women without ovarian stimulation.

Natural cycle IVF produced the world first successful live birth, but slowly this treatment has been replaced by ovarian stimulated cycle IVF, because it has been believed ovarian stimulated cycle IVF will increase the number of available embryos for transfer. Therefore, it directly increases the chance of pregnancy from the treatment cycle. However, ovarian stimulation is always associated with side effects.

Clinical-pregnancy outcome after vitrification of blastocysts produced from in vitro maturation cycles.

This study was conducted to report the clinical-pregnancy outcome after vitrification of the blastocysts produced from in vitro maturation cycles. The survival rate after thawing was 92.0% (92/100).

Survivin acts as anti-apoptotic factor during the development of bovine pre-implantation embryos.

Survivin, an inhibitor of apoptotic protein containing a single baculoviral inhibit apoptotic protein repeat domain, is a bifunctional protein that suppresses apoptosis and regulates cell division. Thus, we used double stranded RNA (dsRNA) interference to manipulate survivin expression in bovine embryos and analyze its role in blocking apoptosis and facilitating development of pre-implantation embryos. In vitro fertilized embryos (1-cell) were injected with survivin dsRNA, and expression of survivin mRNA was evaluated by real-time quantitative RT-PCR.

Effect of gonadotrophin priming on in-vitro maturation of oocytes collected from women at risk of OHSS.

This study examined the effects of human menopausal gonadotrophin (HMG) or human chorionic gonadotrophin (HCG) priming on cumulus-oocyte complex (COC) morphology, oocyte maturation and embryo development in patients undergoing in-vitro maturation (IVM) cycles. The patients were primed with nothing (group 1), low-dose HMG (group 2) or 10,000 IU HCG (group 3) before oocyte retrieval. COC with dispersed cumulus cell appearance was only observed in group 3.

Sterile filtered paraffin oil supports in vitro developmental competence in bovine embryos comparable to co-culture.

Purpose: To investigate whether sterile filtered light paraffin oil (SPO) overlaying is superior to washed light mineral oil (WMO) in supporting the in vitro developmental competence of bovine follicular oocytes. In addition, the effects of the two types of oil overlaying were compared with oil overlaying plus co-culture (CC) on bovine embryo development in vitro.

Methods: Bovine follicular oocytes retrieved from abattoir-derived ovary were in vitro matured, fertilized and cultured in 50 microL drops overlayed with WMO or SPO and were subsequently evaluated for development rates.

Increase in DNA fragmentation and apoptosis-related gene expression in frozen-thawed bovine blastocysts.

Evaluation of apoptosis and expression level of apoptosis-related genes is useful for examining the variation in embryo quality according to environmental change. The objective of this study was to investigate DNA fragmentation and apoptosis-related gene expression patterns in frozen-thawed bovine blastocysts. In vitro produced day 7 blastocysts were frozen by two different vitrification methods (conventional 0.

Optimization of a dilution method for human expanded blastocysts vitrified using EM grids after artificial shrinkage.

Purpose: To verify a more effective dilution method that can be applied to human expanded blastocysts that are vitrified after artificial shrinkage.

Methods: Surplus expanded blastocysts that remained after embryo transfer (ET) in in vitro fertilization (IVF) cycles, were cryopreserved. The blastocysts were vitrified on EM grids following artificial shrinkage.

Fertilization, cleavage and blastocyst development according to the maturation timing of oocytes in in vitro maturation cycles.

Background: This study was to examine the developmental capacity of oocytes collected from an in vitro maturation (IVM) programme according to their maturation time.

Methods: The study included 47 IVM cycles that underwent blastocyst transfer. The patients (n = 38) were primed with 10 000 IU HCG 36 h before their oocyte retrieval.

Correlation between in vitro maturation and expression of LH receptor in cumulus cells of the oocytes collected from PCOS patients in HCG-primed IVM cycles.

Background: The aim of this study was to investigate whether in vitro maturation (IVM) and blastocyst development of oocytes collected following HCG-primed IVM cycles of PCOS patients are correlated with their cumulus cell (CC) patterns and further to investigate mRNA expression of the receptors for FSH, LH and epidermal growth factor (EGF) in the CCs with each pattern.

Methods: Patients who underwent IVM were primed with 10,000 IU of HCG 36 h before oocyte aspiration. The isolated cumulus-oocyte complexes were divided into three groups according to the CC patterns: oocytes with dispersed CCs (group A), oocytes with compacted CCs (group B) and oocytes with sparse CCs (group C).

Pregnancy resulting from transfer of repeat vitrified blastocysts produced by in-vitro matured oocytes in patient with polycystic ovary syndrome.

This report describes a live birth produced from repeat vitrification and thawing of blastocysts derived from in-vitro matured (IVM) oocytes in a woman with polycystic ovarian syndrome. Immature oocyte retrieval was performed on day 12 of her induced menstrual cycle. The patient was administered 10,000 IU of human chorionic gonadotrophin s.

Generation of dopaminergic neurons in vitro from human embryonic stem cells treated with neurotrophic factors.

The aim of this study was to produce dopaminergic neurons in vitro from human embryonic stem (hES) cells following treatment of various neurotrophic factors. MB03 hES cells were induced by retinoic acid (RA) or basic fibroblast growth factor (bFGF), which were further treated with brain derived neurotrophic factor (BDNF) or transforming growth factor (TGF)-alpha in each induction method during neuron differentiation days. At the final differentiation stage (21 days), all treatment groups revealed very similar levels (bFGF, 76-78%; RA, 70-74%) of mature neurons (anti-NF-200) in two induction methods irrespective of the addition of BDNF or TGF-alpha.

Establishment of human embryonic stem cell lines from frozen-thawed blastocysts using STO cell feeder layers.

Background: Recently, human embryonic stem (hES) cells have become very important resources for basic research on cell replacement therapy and other medical applications. The purpose of this study was to test whether pluripotent hES cell lines could be successfully derived from frozen-thawed embryos that were destined to be discarded after 5 years in a routine human IVF-embryo transfer programme and whether an STO cell feeder layer can be used for the culture of hES cells.

Methods: Donated frozen embryos (blastocysts or pronuclear) were thawed, and recovered or in vitro developed blastocysts were immunosurgically treated.

Genetically modified human embryonic stem cells relieve symptomatic motor behavior in a rat model of Parkinson's disease.

Embryonic stem (ES) cells have great potential as a cell source for cell replacement therapy. To investigate the possibility of using ES cells as a carrier of therapeutic gene(s), human ES cells (MB03) were co-transfected with cDNAs coding for tyrosine hydroxylase (TH) and GTP cyclohydrolase I (GTPCH I), then bulk-selected in the presence of neomycin and hygromycin-B. Successful transfection was confirmed by Western immunoblotting and RT-PCR.

Visualization of the metaphase II meiotic spindle in living human oocytes using the Polscope enables the prediction of embryonic developmental competence after ICSI.

Background: Meiotic spindles in living human oocytes can be visualized by the Polscope. This study investigated the relationship between the presence/location of the spindle in metaphase II (MII) oocytes and developmental competence of embryos in vitro.

Methods: The spindles in 626 MII oocytes were examined by the Polscope and divided into six groups (A-F) based on the presence or absence of the spindles and the angle between the spindle and the first polar body.

Pregnancy outcome following transfer of human blastocysts vitrified on electron microscopy grids after induced collapse of the blastocoele.

Background: The purpose of this study was to examine the effects on blastocyst survival and subsequent pregnancy rate of 'artificial shrinkage' (i.e. induced collapse of the blastocoele) before vitrification of human blastocysts.

Successful birth after transfer of blastocysts derived from oocytes of unstimulated woman with regular menstrual cycle after IVM approach.

Purpose: To report a delivery after transfer of blastocysts derived from eggs collected following in vivo HCG priming in a patient with regular menstrual cycles undergoing in vitro maturation (IVM) program.

Methods: A woman had regular menstrual cycle and had experience of ovarian hyperstimulation syndrome (OHSS) during a previous conventional IVF-ET cycle. The patient was primed with 10,000 IU HCG 36 h before egg retrieval.

Ongoing twin pregnancy after vitrification of blastocysts produced by in-vitro matured oocytes retrieved from a woman with polycystic ovary syndrome: Case report.

This case report describes an ongoing pregnancy after cryopreservation of blastocysts produced by in-vitro matured oocytes retrieved from a woman with polycystic ovary syndrome (PCOS). Oocyte retrieval was performed on day 18. The patient was administered 10 000 IU of hCG s.

An improved protocol for dilution of cryoprotectants from vitrified human blastocysts.

Background: The purposes of this study were to compare the survival rate of human blastocysts thawed by two different methods after vitrification using electron microscopic (EM) grids, and to report the successful pregnancies and live births resulting from the transfer of blastocysts that had survived the established thawing method.

Methods: The blastocysts produced from three pronuclei zygotes in IVF cycles were vitrified on an EM grid. After dilution of cryoprotectant by either a 6- or 2-step method, the survival rate of the blastocysts was compared.

Blastocyst development and pregnancies after IVF of mature oocytes retrieved from unstimulated patients with PCOS after in-vivo HCG priming.

A major side-effect of controlled ovarian stimulation (COS) in patients with polycystic ovarian syndrome (PCOS) is the risk of ovarian hyperstimulation syndrome (OHSS). In-vitro maturation (IVM) of immature oocytes represents a potential alternative for the fertility treatment of these patients. Two patients at high risk of OHSS were primed with 10,000 IU HCG 36 h before oocyte retrieval.