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Blastocyst culture

Hiroshi Tajimi, Tsugumi Yamazaki, Syoko Oike, Tatsuyuki Yoshida, Konosuke Okada, Masashige Kuwayama, Hitoshi Ushijima
This study was conducted to examine the utility of vitrification for bovine embryos with low-quality grade, and simple cryoprotectants dilution method for practitioners. In Experiment 1, survival of frozen embryos was compared with that of vitrified embryos using minimum volume cooling (MVC). Then, vitrified embryos were used to confirm the optimum sucrose concentration in Experiment 2. The survival rates of embryos that had been vitrified following diluted cryoprotectants with the one-step in-straw method were compared with those of fresh control embryos in Experiment 3...
May 17, 2018: Animal Science Journal, Nihon Chikusan Gakkaihō
Fuminori Tanihara, Maki Hirata, Nguyen Thi Nhien, Takayuki Hirano, Toshiki Kunihara, Takeshige Otoi
The value of laboratory and genetically-modified pigs is becoming increasingly clear; however, their in vitro development remains inefficient. Trans-ferulic acid (trans-FA) is an aromatic compound that is abundant in plant cell walls, and which exhibits antioxidant effects in vitro. Trans-FA is known to improve sperm viability and motility; however, its effects on porcine oocytes are unknown. Our aim was to investigate the effects of trans-FA supplementation during in vitro maturation on the meiotic and developmental competence of porcine oocytes...
May 16, 2018: Journal of Veterinary Medical Science
Suo Li, Qing Guo, Yu-Meng Wang, Zi-Yue Li, Jin-Dan Kang, Xi-Jun Yin, Xin Zheng
Glycine, a component of glutathione (GSH), plays an important role in protection from reactive oxygen species (ROS) and inhibition of apoptosis. The aim of this study was to determine the effect of glycine on in vitro maturation (IVM) of porcine oocytes and their developmental competence after parthenogenetic activation (PA). We examined nuclear maturation, ROS levels, apoptosis, mitochondrial membrane potential (ΔΨm), and ATP concentration, as well as the expression of several genes related to oocyte maturation and development...
May 12, 2018: Journal of Animal Science
Chuti Laowtammathron, Pimjai Chingsuwanrote, Roungsin Choavaratana, Suphadtra Phornwilardsiri, Ketsara Sitthirit, Chidchanok Kaewjunun, Orawan Makemaharn, Papussorn Terbto, Supaporn Waeteekul, Chanchao Lorthongpanich, Yaowalak U-Pratya, Pimonwan Srisook, Pakpoom Kheolamai, Surapol Issaragrisil
BACKGROUND: Due to their extensive self-renewal and multilineage differentiation capacity, human embryonic stem cells (hESCs) have great potential for studying developmental biology, disease modeling, and developing cell replacement therapy. The first hESC line was generated in 1998 by culturing inner cell mass (ICM) cells isolated from human blastocysts using an immunosurgery technique. Since then, many techniques including mechanical ICM isolation, laser dissection, and whole embryo culture have been used to derive hESC lines...
May 11, 2018: Stem Cell Research & Therapy
Valeriy Kuznyetsov, Svetlana Madjunkova, Ran Antes, Rina Abramov, Gelareh Motamedi, Zenon Ibarrientos, Clifford Librach
OBJECTIVE: To assess whether embryonic DNA isolated from blastocyst culture conditioned medium (BCCM) combined with blastocoel fluid (BF) could be used for blastocyst stage non-invasive preimplantation genetic testing for chromosomal aneuploidy (non-invasive preimplantation genetic screening, NIPGS). PATIENTS: 47 embryos from 35 patients undergoing IVF. INTERVENTIONS: DNA analysis of combined BCCM plus BF in comparison with trophectoderm (TE) biopsy and/or whole blastocyst (WB)using next generation sequencing (NGS)...
2018: PloS One
Kanokwan Srirattana, Justin C St John
We generated cattle embryos using mitochondrial supplementation and somatic cell nuclear transfer (SCNT), named miNT, to determine how additional mitochondrial DNA (mtDNA) modulates the nuclear genome. To eliminate any confounding effects from somatic cell mtDNA in intraspecies SCNT, donor cell mtDNA was depleted prior to embryo production. Additional oocyte mtDNA did not affect embryo development rates but increased mtDNA copy number in blastocyst stage embryos. Moreover, miNT-derived blastocysts had different gene expression profiles when compared with SCNT-derived blastocysts...
May 8, 2018: Scientific Reports
E J Hoekman, K E Tucker, C Am Jansen, C G Hilders
BACKGROUND: Oocyte vitrification is important for fertility preservation. There are debates, however, surrounding the components of the procedure itself. OBJECTIVE: before starting a vitrification program, we decided to (1) determine if blastocysts derived from previously frozen mouse embryos would be suitable to practice vitrification and (2) to analyze the factors that contributed to improving our "learning curve". MATERIALS AND METHODS: 58 expanded blastocysts cultured from commercially available frozen 1-cell mouse embryos (B6D2F1 x B6C3F1) were used...
March 2018: Cryo Letters
S S Sahraei, M Shahhoseini, B Movaghar
BACKGROUND: OBJECTIVE: This paper reports results from studies on modifications of histone marks in transcriptional regulatory regions of H19, Igf2 and Mest genes in vitrified-warmed and cultured mouse embryos. MATERIALS AND METHODS: Non vitrified and vitrified-warmed 8-cell embryos were cultured in Ham's F10+10% BSA into blastocyst stage in two experimental groups. Gene expression level was assessed by quantitative PCR and histone modifications were evaluated by ChIP assay method...
March 2018: Cryo Letters
J J Dai, J H Yang, S S Zhang, Y F Niu, Y N Chen, C F Wu, D F Zhang
  The survival of porcine oocytes is still very low after cryopreservation. OBJECTIVE: To investigate whether and when the mitochondrial function of vitrified porcine oocytes could be recovered post-thaw. MATERIALS AND METHODS: Mitochondrial potential, ROS level, ATP content, apoptotic rate, caspase activity, and parthenogenetics developmental ability of thawed porcine oocytes were measured after culture in vitro for 0, 1, 2 or 4 h. RESULTS: Mitochondrial potential after 2 h and 4 h post-thaw culture were 1...
January 2018: Cryo Letters
V H Do, S Watson, S Catt, A W Taylor-Robinson
  BACKGROUND: In order to thaw slow-cooled bovine embryos it is standard practice to draw out permeating cryoprotectants by passing embryos through successively decreasing osmotic solutions. However, recently it has been suggested that sucrose may not be needed in the warming media. OBJECTIVE: The aim of this experiment was to compare the effect of warming media prepared with or without the inclusion of sucrose on the survival and hatching capacity of vitrified in vitro-derived bovine embryos...
September 2017: Cryo Letters
Toshinori Oikawa, Tomoko Itahashi, Risa Yajima, Takashi Numabe
Intracytoplasmic sperm injection (ICSI) was expected to enable more efficient use of sperm from sires with preferable genetic traits and result in a generation containing a larger number of offspring with superior genetic characteristics in livestock. However, the efficiency of the early development of embryos produced by ICSI is still far from satisfactory in cattle. The present study aimed to investigate the effects of the treatment of cryopreserved sperm with glutathione (GSH) on the early development of embryos produced by ICSI in Japanese Black cattle...
May 7, 2018: Journal of Reproduction and Development
Sun-Hee Lee, Chan Woo Park, Yong-Pil Cheon, Chun Kyu Lim
PURPOSE: Recent studies have shown that improved clinical outcomes can be achieved by transferring blastocysts rather than cleavage-stage embryos. However, blastocyst transfer is not performed in all patients. The aim of this study was to compare clinical outcomes of intracytoplasmic sperm injection (ICSI) cycles using testicular sperm (TE) with those of ICSI cycles using ejaculated sperm (EJ). METHODS: ICSI was performed using EJ in 141 cycles and TE in 37 cycles...
May 5, 2018: Journal of Assisted Reproduction and Genetics
Danilo Cimadomo, C Scarica, R Maggiulli, G Orlando, D Soscia, L Albricci, S Romano, F Sanges, F M Ubaldi, L Rienzi
PURPOSE: To assess whether continuous embryo culture involves better embryological and/or clinical outcomes than sequential. METHODS: Prospective study at a private IVF center. All consecutive IVF cycles (September 2013-2015) fulfilling the inclusion criteria underwent embryo culture in either Continuous-Single-Culture-Media (CSCM, n = 972) or sequential media (Quinn's Advantage, n = 514), respectively. ICSI, blastocyst culture in either standard (MINC) or undisturbed (Embryoscope) incubation, transfer (until September 2016), and pregnancy follow-up (until September 2017) were performed...
May 3, 2018: Journal of Assisted Reproduction and Genetics
Shichao Liu, Gerelchimeg Bou, Jianchao Zhao, Shimeng Guo, Jia Guo, Xiaogang Weng, Zhi Yin, Zhonghua Liu
Upregulation of Cdx2 expression in outer cells is a key event responsible for cell lineage segregation between inner cell mass (ICM) and trophoderm (TE) in mouse morula stage embryos. In TE cells, polarization can regulate Hippo and ROCK signaling to induce the nuclear location of YAP, which has been demonstrated to further induce the expression of Cdx2. However, we found that CDX2 expression could not be detected in the outer cells of porcine morula stage embryos and could be found in only some TE cells at the early blastocyst stage...
May 2, 2018: Molecular Reproduction and Development
Ying Zhang, Pengxiang Qu, Xiaonan Ma, Fang Qiao, Yefei Ma, Suzhu Qing, Yong Zhang, Yongsheng Wang, Wei Cui
Serum starvation is a routine protocol for synchronizing nuclear donor cells to G0/G1 phase during somatic cell nuclear transfer (SCNT). However, abrupt serum deprivation can cause serious stress to the cells cultured in vitro, which might result in endoplasmic reticulum (ER) stress, chromosome damage, and finally reduce the success rate of SCNT. In the present study, the effects of tauroursodeoxycholic acid (TUDCA), an effective ER stress-relieving drug, on the nuclear donor cells under serum deprivation condition as well as following SCNT procedures were first assessed in the bovine...
2018: PloS One
Erif Maha Nugraha Setyawan, Hyun Ju Oh, Min Jung Kim, Geon A Kim, Seok Hee Lee, Yoo Bin Choi, Kihae Ra, Byeong Chun Lee
The paracrine interactions between cumulus-oocyte complexes (COCs) and follicular somatic cells during in vitro maturation (IVM) were investigated. To optimize IVM conditions, many studies have applied exogenous growth factors and cell feeding/co-culture systems using various cell types to replicate the natural follicular microenvironment during IVM. A potential candidate as cell feeders is adipose-derived stem cells (ASCs) which secrete high levels of growth factors that have roles in oocyte maturation. However, the cell donor's age should be considered because biological aging also occurs in stem cells...
December 12, 2017: Theriogenology
Jolanta Opiela, Bülent Bülbül, Joanna Romanek
The objective of the present study was to evaluate the effect of porcine mesenchymal stem cells (MSCs) secreted factors on bovine in vitro embryo development using MSCs in different culture systems: SOF medium, SOF medium conditioned by MSCs in monolayer, and in reverse drop and by embryo culture in coculture with MSCs. Statistically highly significant differences were noted between the number of blastocysts derived cultures in all tested culture systems. The in vitro culture in SOF turned out to be the most optimal...
April 30, 2018: Animal Biotechnology
Saghar Saeedabadi, Amir Hossein Abazari-Kia, Hoda Rajabi, Kazem Parivar, Mohammad Salehi
Background: DNA methylation is one the epigenetic mechanisms, which is critically involved in gene expression. This phenomenon is mediated by DNA methyl-transferases and is affected by environmental stress, including in vitro maturation (IVM) of oocytes. Melatonin, as an antioxidant, may theoretically be involved in epigenetic regulation via reductions of reactive oxygen species. This study was performed to investigate DNA methylation and the possibility of goat oocyte development after treatment with different concentrations of melatonin...
July 2018: International Journal of Fertility & Sterility
Hao Jiang, Shuang Liang, Xue-Rui Yao, Yong-Xun Jin, Xing-Hui Shen, Bao Yuan, Jia-Bao Zhang, Nam-Hyung Kim
Laminarin (LMA), a β-glucan mixture with good biocompatibility, improves the growth performance and immune response when used as food additives and nutraceuticals. The aim of the present research was to explore the effects of LMA on porcine early stage embryo development, as well as the underlying mechanisms. The results showed that the developmental competence of porcine early stage embryos was dramatically improved after LMA supplementation during the in vitro culture period. The presence of 20 μg/mL LMA during the in vitro culture period significantly improved cleavage rate, blastocyst formation rates, hatching rate, and total cell number in the blastocyst compared to that in the control group...
April 23, 2018: Theriogenology
Casey C Read, Gabby Willhelm, Paul W Dyce
Gap junctional coupling between cumulus cells is required for oocytes to reach developmental competence. Multiple connexins, which form these gap junctions, have been found within the ovarian follicles of several species including bovine. The aim of this study was to determine the role of connexin 43 (CX43) and its relationship to embryo development, following in vitro fertilization (IVF). Cumulus-oocyte-complexes (COCs) were obtained from abattoir sourced, mixed breed, bovine ovaries. COCs were isolated from follicles ranging from 2-5mm in size, representing the preselected follicle pool...
April 26, 2018: Molecular Reproduction and Development
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