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Hu Y.,Vivere Health
Fertility and Sterility | Year: 2016

Objective To report a live birth of twins from autologous zona-free oocytes. Design Case report. Setting Reproductive endocrinology and infertility private practice and ambulatory in vitro fertilization (IVF) center. Patient(s) A 34-year-old woman, gravida 0, with 100% zona-free oocytes. Intervention(s) IVF with intracytoplasmic sperm injection (ICSI), blastocyst culture, and fresh embryo transfer. Main Outcome Measure(s) Fertilization, blastocyst development, and live birth. Result(s) A 34-year-old woman, gravida 0, conceived through IVF using autologous zona-free oocytes. The 11 retrieved oocytes were zona-free, from which eight were inseminated with ICSI; two embryos were transferred at morula and blastocyst stage, resulting in a twin pregnancy delivered at an estimated the gestational age of 37 weeks and 1 day. Conclusion(s) Patients with the rare condition of 100% zona-free oocytes maintain the potential for pregnancy after careful micromanipulation of the oocytes. Caution is recommended on the number of embryos selected for transfer to reduce the risk of multiple gestation. © 2016 American Society for Reproductive Medicine, Published by Elsevier Inc.


He W.,Guangzhou University | Sun X.,Guangzhou University | Liu L.,pacgenomics | Li M.,pacgenomics | And 3 more authors.
PLoS ONE | Year: 2014

Chromosomal anomalies in human embryos produced by in vitro fertilization are very common, which include numerical (aneuploidy) and structural (deletion, duplication or others) anomalies. Our previous study indicated that chromosomal deletion(s) is the most common structural anomaly accounting for approximately 8% of euploid blastocysts. It is still unknown if these deletions in human euploid blastocysts have clinical significance. In this study, we analyzed 15 previously diagnosed euploid blastocysts that had chromosomal deletion(s) using Agilent oligonucleotide DNA microarray platform and localized the gene location in each deletion. Then, we used OMIM gene map and phenotype database to investigate if these deletions are related with some important genes that cause genetic diseases, especially developmental delay or intellectual disability. As results, we found that the detectable chromosomal deletion size with Agilent microarray is above 2.38 Mb, while the deletions observed in human blastocysts are between 11.6 to 103 Mb. With OMIM gene map and phenotype database information, we found that deletions can result in loss of 81-464 genes. Out of these genes, 34-149 genes are related with known genetic problems. Furthermore, we found that 5 out of 15 samples lost genes in the deleted region, which were related to developmental delay and/or intellectual disability. In conclusion, our data indicates that all human euploid blastocysts with chromosomal deletion(s) are abnormal and transfer of these embryos may cause birth defects and/or developmental and intellectual disabilities. Therefore, the embryos with chromosomal deletion revealed by DNA microarray should not be transferred to the patients, or further gene map and/or phenotype seeking is necessary before making a final decision. Copyright: © 2014 Tondeleir et al.


Zheng H.,Guangzhou University | Jin H.,pacgenomics | Liu L.,pacgenomics | Liu J.,Guangzhou University | Wang W.-H.,Vivere Health
Molecular Cytogenetics | Year: 2015

Background: Aneuploidy is a leading cause of repeat implantation failure and recurrent miscarriages. Preimplantation genetic screening (PGS) enables the assessment of the numeral and structural chromosomal errors of embryos before transfer in patients undergoing in vitro fertilization. Array comparative genomic hybridization (aCGH) has been demonstrated to be an accurate PGS method and in present thought to be the gold standard, but new technologies, such as next-generation sequencing (NGS), continue to emerge. Validation of the new comprehensive NGS-based 24-chromosome aneuploidy screening technology is still needed to determine the preclinical accuracy before it might be considered as an alternative method for human PGS. Results: In the present study, 43 human trophectoderm (TE) biopsy samples and 5 cytogenetically characterized cell lines (Coriell Cell Repositories) were tested. The same whole genome amplified product of each sample was blindly assessed with Veriseq NGS and Agilent aCGH to identify the aneuploidy status. The result showed that the NGS identified all abnormalities identified in aCGH including the numeral chromosomal abnormalities (again or loss) in the embryo samples and the structural (partial deletion and duplication) in the Coriell cell lines. Both technologies can identify a segmental imbalance as small as 1.8 Mb in size. Among the 41 TE samples with abnormal karyotypes in this study, eight (19.5 %) samples presented as multiple chromosome abnormalities. The abnormalities occurred to almost all chromosomes, except chromosome 6, 7, 17 and Y chromosome. Conclusions: Given its reliability and high level of consistency with an established aCGH methodology, NGS has demonstrated a robust high-throughput methodology ready for extensive clinical application in reproductive medicine, with potential advantages of reduced costs and enhanced precision. Then, a randomized controlled clinical trial confirming its clinical effectiveness is advisable to obtain a larger sequencing dataset and more evidence for the extensive use of NGS-based PGS. © 2015 Zheng et al.


PubMed | Vivere Health
Type: Journal Article | Journal: Fertility and sterility | Year: 2016

To report a live birth of twins from autologous zona-free oocytes.Case report.Reproductive endocrinology and infertility private practice and ambulatory invitro fertilization (IVF) center.A 34-year-old woman, gravida 0, with 100% zona-free oocytes.IVF with intracytoplasmic sperm injection (ICSI), blastocyst culture, and fresh embryo transfer.Fertilization, blastocyst development, and live birth.A 34-year-old woman, gravida 0, conceived through IVF using autologous zona-free oocytes. The 11 retrieved oocytes were zona-free, from which eight were inseminated with ICSI; two embryos were transferred at morula and blastocyst stage, resulting in a twin pregnancy delivered at an estimated the gestational age of 37weeks and 1day.Patients with the rare condition of 100% zona-free oocytes maintain the potential for pregnancy after careful micromanipulation of the oocytes. Caution is recommended on the number of embryos selected for transfer to reduce the risk of multiple gestation.


PubMed | Guangzhou University, Vivere Health and pacgenomics
Type: | Journal: Molecular cytogenetics | Year: 2015

Aneuploidy is a leading cause of repeat implantation failure and recurrent miscarriages. Preimplantation genetic screening (PGS) enables the assessment of the numeral and structural chromosomal errors of embryos before transfer in patients undergoing in vitro fertilization. Array comparative genomic hybridization (aCGH) has been demonstrated to be an accurate PGS method and in present thought to be the gold standard, but new technologies, such as next-generation sequencing (NGS), continue to emerge. Validation of the new comprehensive NGS-based 24-chromosome aneuploidy screening technology is still needed to determine the preclinical accuracy before it might be considered as an alternative method for human PGS.In the present study, 43 human trophectoderm (TE) biopsy samples and 5 cytogenetically characterized cell lines (Coriell Cell Repositories) were tested. The same whole genome amplified product of each sample was blindly assessed with Veriseq NGS and Agilent aCGH to identify the aneuploidy status. The result showed that the NGS identified all abnormalities identified in aCGH including the numeral chromosomal abnormalities (again or loss) in the embryo samples and the structural (partial deletion and duplication) in the Coriell cell lines. Both technologies can identify a segmental imbalance as small as 1.8Mb in size. Among the 41 TE samples with abnormal karyotypes in this study, eight (19.5%) samples presented as multiple chromosome abnormalities. The abnormalities occurred to almost all chromosomes, except chromosome 6, 7, 17 and Y chromosome.Given its reliability and high level of consistency with an established aCGH methodology, NGS has demonstrated a robust high-throughput methodology ready for extensive clinical application in reproductive medicine, with potential advantages of reduced costs and enhanced precision. Then, a randomized controlled clinical trial confirming its clinical effectiveness is advisable to obtain a larger sequencing dataset and more evidence for the extensive use of NGS-based PGS.


Deng A.,Vivere Health | Wang W.-H.,Houston Fertility Institute | Wang W.-H.,Changsha Hospital for Maternal and Child Health Care
Molecular Cytogenetics | Year: 2015

Background: Increased embryo implantation rates were reported after transfer of euploid embryos selected by preimplantation genetic screening (PGS). Egg cryopreservation by vitrification has become one of the most important assisted human reproduction technologies. Although reports indicate that development and implantation of human embryos derived from frozen donor eggs are comparative to fresh eggs, it is still unknown whether egg vitrification increases chromosomal abnormalities in eggs, which in turn causes formation of embryonic aneuploidy. Therefore, in this study, we evaluated the aneuploidy formation in the blastocysts derived from frozen donor eggs and also evaluated the efficiency of egg vitrification as an advanced technology for egg cryopreservation. Results: In this study, donated human eggs from young women were cryopreserved by vitrification and PGS was performed in the resulted blastocysts by DNA microarray. A total of 764 frozen eggs from 75 egg thawing cycles were warmed and 38 blastocysts were biopsied for PGS before embryo transfer. A 97.1% of egg survival rate was obtained and 59.1% of embryos developed to blastocyst stage. After biopsy and PGS, it was found that 84.2% of blastocysts were euploid and 15.8% were aneuploid. Aneuploidy rates varied among donors. Transfers of blastocysts without PGS resulted in higher clinical pregnancy and implantation rates as compared with transfer of blastocysts with PGS. Conclusions: Although the overall aneuploidy rate was low in the blastocysts derived from frozen donor eggs, high aneuploidy rates were observed in the embryos resulting from some donated eggs. Clinical pregnancy rate was not improved by PGS of embryos resulting from donor eggs, indicating that PGS may not be necessary for embryos derived from donor eggs in most cases. © 2015 Deng and Wang; licensee BioMed Central.


PubMed | Changsha Hospital for Maternal and Child Health Care and Vivere Health
Type: | Journal: Molecular cytogenetics | Year: 2015

Increased embryo implantation rates were reported after transfer of euploid embryos selected by preimplantation genetic screening (PGS). Egg cryopreservation by vitrification has become one of the most important assisted human reproduction technologies. Although reports indicate that development and implantation of human embryos derived from frozen donor eggs are comparative to fresh eggs, it is still unknown whether egg vitrification increases chromosomal abnormalities in eggs, which in turn causes formation of embryonic aneuploidy. Therefore, in this study, we evaluated the aneuploidy formation in the blastocysts derived from frozen donor eggs and also evaluated the efficiency of egg vitrification as an advanced technology for egg cryopreservation.In this study, donated human eggs from young women were cryopreserved by vitrification and PGS was performed in the resulted blastocysts by DNA microarray. A total of 764 frozen eggs from 75 egg thawing cycles were warmed and 38 blastocysts were biopsied for PGS before embryo transfer. A 97.1% of egg survival rate was obtained and 59.1% of embryos developed to blastocyst stage. After biopsy and PGS, it was found that 84.2% of blastocysts were euploid and 15.8% were aneuploid. Aneuploidy rates varied among donors. Transfers of blastocysts without PGS resulted in higher clinical pregnancy and implantation rates as compared with transfer of blastocysts with PGS.Although the overall aneuploidy rate was low in the blastocysts derived from frozen donor eggs, high aneuploidy rates were observed in the embryos resulting from some donated eggs. Clinical pregnancy rate was not improved by PGS of embryos resulting from donor eggs, indicating that PGS may not be necessary for embryos derived from donor eggs in most cases.


Wang C.,Purdue University | Wang C.,Vivere Health | Zhang L.,Purdue University | Jaeger L.A.,Purdue University | Machaty Z.,Purdue University
Biology of Reproduction | Year: 2015

The role of store-operated Ca2+ entry (SOCE) in the maintenance of sperm-induced Ca2+ oscillations was investigated in porcine eggs. We found that 10 μM gadolinium (Gd3+), which is known to inhibit SOCE, blocked Ca2+ entry that was triggered by thapsigargin-induced store depletion and also caused an abrupt cessation of the fertilization Ca2+ signal. In a similar manner 3,5-bis(trifluoromethyl)pyrazole 2 (20 lM), and tetrapandin-2 (10 μM), potent SOCE inhibitors, also blocked thapsigargin-stimulated Ca2+ entry and disrupted the Ca2+ oscillations after sperm-egg fusion. The downregulation of Stim1 or Orai1 in the eggs did not alter the Ca2+ content of the intracellular stores, whereas co-overexpression of these proteins led to the generation of irregular Ca2+ transients after fertilization that stopped prematurely. We also found that thapsigargin completely emptied the endoplasmic reticulum, and that the series of Ca2+ transients stopped abruptly after the addition of thapsigargin to the fertilized eggs, indicating that the proper reloading of the intracellular stores is a prerequisite for the maintenance of the Ca2+ oscillations. These data strengthen our previous findings that in porcine eggs SOCE is a major signaling cascade that is responsible for sustaining the repetitive Ca2+ signal at fertilization. © 2015 by the Society for the Study of Reproduction, Inc.


Herrick J.R.,National Foundation for Fertility Research | Wang C.,Purdue University | Wang C.,Vivere Health | Machaty Z.,Purdue University
Reproduction, Fertility and Development | Year: 2016

Embryos produced from vitrified feline oocytes have resulted in pregnancies, but the efficiency of oocyte vitrification in cats is still low. Our objectives were to evaluate the effects of exposing feline oocytes to ethylene glycol (EG), propanediol (PrOH) and dimethyl sulfoxide (DMSO) on changes in intracellular free-calcium concentrations ([Ca2+]i), the time needed for enzymatic digestion of the zona pellucida (ZP), the incidence of parthenogenetic activation and degeneration and embryonic development following in vitro fertilisation (IVF). All of the chemicals tested altered [Ca2+]i, but changes in [Ca2+]i, resistance of the ZP to enzymatic digestion and the incidence of parthenogenetic activation (<5% for all treatments) were not affected (P>0.05) by extracellular Ca2+. Exposure to EG (>44.1%) and DMSO (19.7%) increased (P<0.05) oocyte degeneration compared with control oocytes and oocytes exposed to PrOH (≤2.5%). Following exposure to a combination of PrOH and DMSO (10% v/v each), blastocyst development (per cleaved embryo; 52.1%) was similar (P>0.05) to control oocytes (64.4%). When oocytes were vitrified with PrOH and DMSO, 28.3% of surviving (intact plasma membrane) oocytes cleaved following IVF, but no blastocyst developed. When a non-permeating cryoprotectant (galactose, 0.25M) was added to the vitrification medium, 47.7% of surviving oocytes cleaved and 14.3% developed to the blastocyst stage. © CSIRO 2016.


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