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Blastocyst culture

G Galeati, E Giaretta, A Zannoni, D Bucci, C Tamanini, M Forni, M Spinaci
Oxidative stress caused from in vitro culture contributes to inadequate oocyte maturation which leads to a poor embryo development. Therefore, it is important to protect oocytes and embryos against oxidative stress. This study was aimed at evaluating the effect of Embelin (2,5-dihydroxy-3-undecyl-1,4-benzoquinone), an antioxidant with various pharmacologic activities, on nuclear and cytoplasmic maturation of pig oocytes as well as on steroidogenesis of cumulus cells (CCs). Another objective was to determine the influence of Embelin on developmental competence of pig oocytes as well as the expression levels of three key genes (Nanog, Sox2 and Oct4) involved in the control of pluripotency in parthenogenetically activated embryos...
August 2016: Journal of Physiology and Pharmacology: An Official Journal of the Polish Physiological Society
Jae Gyu Yoo, Byeong-Woo Kim, Mi-Rung Park, Deug-Nam Kwon, Yun-Jung Choi, Teak-Soon Shin, Byung-Wook Cho, Jakyeom Seo, Jin-Hoi Kim, Seong-Keun Cho
Objective: The present study investigates pre- and post-implantation developmental competence of nuclear-transferred porcine embryos derived from male and female fetal fibroblasts. Methods: Male and female fetal fibroblasts were transferred to in vitro-matured enucleated oocytes and in vitro and in vivo developmental competence of reconstructed embryos was investigated. And, a total of 6,789 female fibroblast nuclear-transferred embryos were surgically transferred into 41 surrogate gilts and 4,746 male fibroblast nuclear-transferred embryos were surgically transferred into 25 surrogate gilts...
October 20, 2016: Asian-Australasian Journal of Animal Sciences
J Wu, A Platero Luengo, M A Gil, K Suzuki, C Cuello, M Morales Valencia, I Parrilla, C A Martinez, A Nohalez, J Roca, E A Martinez, J C Izpisua Belmonte
More than eighteen years have passed since the first derivation of human embryonic stem cells (ESCs), but their clinical use is still met with several challenges, such as ethical concerns regarding the need of human embryos, tissue rejection after transplantation and tumour formation. The generation of human induced pluripotent stem cells (iPSCs) enables the access to patient-derived pluripotent stem cells (PSCs) and opens the door for personalized medicine as tissues/organs can potentially be generated from the same genetic background as the patient recipients, thus avoiding immune rejections or complication of immunosuppression strategies...
October 2016: Reproduction in Domestic Animals, Zuchthygiene
Tong Zhang, Chunqiang Zhang, Xiaomei Fan, Ruilan Li, Jiaxin Zhang
C-type natriuretic peptide (CNP) has been considered as a physiological meiotic inhibitor that stimulates the cGMP production by cumulus cell natriuretic peptide receptor 2 (NPR2), which inhibits oocyte phosphodiesterase type 3 activity and increases cAMP. In this study, we explored the effect of CNP pretreatment on the in vitro maturation (IVM) of bovine oocytes by examining changes in cleavage rate, blastocyst formation, mitochondrial DNA (mtDNA) copy number, reactive oxygen species (ROS) level, glutathione (GSH) content, and redox state...
October 19, 2016: In Vitro Cellular & Developmental Biology. Animal
A L S Guimarães, S A Pereira, M N Diógenes, M A N Dode
The aim of this study was to evaluate the effect of adding a combination of insulin, transferrin and selenium (ITS) and l-ascorbic acid (AA) during in vitro maturation (IVM) and in vitro culture (IVC) on in vitro embryo production. To verify the effect of the supplements, cleavage and blastocyst rates, embryo size and total cell number were performed. Embryonic development data, embryo size categorization and kinetics of maturation were analyzed by chi-squared test, while the total cell number was analyzed by a Kruskal-Wallis test (P < 0...
October 17, 2016: Zygote: the Biology of Gametes and Early Embryos
Xiaofeng Zheng, Pengyi Yang, Brad Lackford, Brian D Bennett, Li Wang, Hui Li, Yu Wang, Yiliang Miao, Julie F Foley, David C Fargo, Ying Jin, Carmen J Williams, Raja Jothi, Guang Hu
Poly(A) tail length and mRNA deadenylation play important roles in gene regulation. However, how they regulate embryonic development and pluripotent cell fate is not fully understood. Here we present evidence that CNOT3-dependent mRNA deadenylation governs the pluripotent state. We show that CNOT3, a component of the Ccr4-Not deadenylase complex, is required for mouse epiblast maintenance. It is highly expressed in blastocysts and its deletion leads to peri-implantation lethality. The epiblast cells in Cnot3 deletion embryos are quickly lost during diapause and fail to outgrow in culture...
October 8, 2016: Stem Cell Reports
Kristin M Whitworth, Joshua A Benne, Lee D Spate, Stephanie L Murphy, Melissa S Samuel, Clifton N Murphy, Jürgen A Richt, Eric Walters, Randall S Prather, Kevin D Wells
The CRISPR/Cas9 genome editing tool has increased the efficiency of creating genetically modified pigs for use as biomedical or agricultural models. The objectives were to determine if DNA editing resulted in a delay in development to the blastocyst stage or in a skewing of the sex ratio. Six DNA templates (gBlocks) that were designed to express guide RNAs that target the transmembrane protease, serine S1, member 2 (TMPRSS2) gene were in vitro transcribed. Pairs of CRISPR guide RNAs that flanked the start codon and polyadenylated Cas9 were co-injected into the cytoplasm of zygotes and cultured in vitro to the blastocyst stage...
October 15, 2016: Transgenic Research
Konstantinos A Economou, Dimitra Christopikou, Erika Tsorva, Stephen Davies, Minas Mastrominas, Haris Cazlaris, Michael Koutsilieris, Panagoula Angelogianni, Dimitris Loutradis
PURPOSE: Artificial oocyte activation using calcium ionophores and enhancement of embryonic developmental potential by the granulocyte-macrophage colony-stimulating factor (GM-CSF) have already been reported. In this study, we evaluated the synergistic effect of these two methods on aged human unfertilized oocytes after intracytoplasmic sperm injection (ICSI). Then, we cultured the resulting embryos to the blastocyst stage and screened them for chromosomal abnormalities, to assess the safety of this protocol...
October 14, 2016: Journal of Assisted Reproduction and Genetics
Diting Qian, Zhiling Li, Yuting Zhang, Yue Huang, Que Wu, Gaizhen Ru, Man Chen, Bin Wang
Our study aimed to develop embryo models to evaluate the impact of oxidative stress on embryo development. Mouse zygotes, which stayed at G1 phase, were treated with prepared culture medium (containing 0.00, 0.01, 0.02, 0.03, 0.04, 0.05, or 0.1 mM hydrogen peroxide (H2O2)) for 30 min in experiment 1. The dose-effects of H2O2 on embryo development were investigated via comparisons of the formation rate at each stage (2- and 4-cell embryos and blastocysts). Experiment 2 was carried out to compare behaviors of embryos in a mild oxidative-stressed status (0...
2016: Oxidative Medicine and Cellular Longevity
E Dietzel, J Floehr, E Van de Leur, R Weiskirchen, W Jahnen-Dechent
STUDY QUESTION: Does fetuin-B inhibit premature zona pellucida (ZP) hardening in mouse oocytes in vitro and thus increase IVF rate? SUMMARY ANSWER: Supplementation of oocyte in vitro maturation (IVM) media with recombinant mouse fetuin-B (rmFetuB) increased fertilization rate without affecting mouse embryo development into blastocysts. WHAT IS KNOWN ALREADY: Mice deficient in fetuin-B are infertile owing to premature ZP hardening. Premature ZP hardening also occurs during oocyte IVM leading to decreased fertilization rate...
October 12, 2016: Molecular Human Reproduction
Yukou Sugano, Manami Yazawa, Sachio Takino, Sueo Niimura, Hideaki Yamashiro
The objective of this study was to classify spindle and first polar body (PB1) chromosome images in ovulated mouse oocytes over time to predict the developmental competence of metaphase II (MII) oocytes. Oocytes were collected at 12, 15, 20, and 25 h after human chorionic gonadotropin (hCG) injection, and stained for spindle tubulin, chromosomes, and PB1 chromosomes. MII spindle morphology was classified as tapered type or barrel type and PB1 chromosomes were categorized as aggregated, separated, dot, or collapsed...
October 13, 2016: Zygote: the Biology of Gametes and Early Embryos
W P Martins, C O Nastri, L Rienzi, S Z van der Poel, C Gracia, C Racowsky
OBJECTIVES: Blastocyst transfer could be advantageous because it allows a more physiologic timing of exposure of the embryo to the uterine environment and permit embryo self-selection after activation of the embryonic genome on day 3. Conversely, the in vitro environment is probably inferior to that provided in vivo and in vitro culture beyond embryonic genomic activation could potentially harm embryos. Our objective was to identify, appraise and summarize the available evidence comparing the effectiveness of blastocyst and cleavage stage embryo transfer...
October 12, 2016: Ultrasound in Obstetrics & Gynecology
Dowglish F Chaves, Iana S Campelo, Mirelly M A S Silva, Maajid H Bhat, Darcio I A Teixeira, Luciana M Melo, Joanna M G Souza-Fabjan, Pascal Mermillod, Vicente J F Freitas
The aim of this study was to evaluate the use of antifreeze protein type III (AFP III) into vitrification medium on meiotic spindle morphology of in vitro matured bovine oocytes as well as the fertilization and blastocyst rates. Mature cumulus-oocyte complexes (COC) were distributed in four groups: control (untreated), vitrified without supplementation (AFP0) or supplemented with 500 (AFP500) or 1000 ng/mL (AFP1000) into vitrification solutions. Samples from each group were used to analyze the organization of meiotic spindle by confocal microscopy and the remaining COC were submitted to in vitro fertilization and culture for eight days...
October 8, 2016: Cryobiology
Kindra Rader, Young-Ho Choi, Katrin Hinrichs
Intracytoplasmic sperm injection is becoming a common clinical procedure in the horse, but little information is available on techniques for its performance. Each laboratory uses different procedures and different media for the steps involved with in vitro embryo production. This article outlines the procedures used in the Clinical Equine Intracytoplasmic Sperm Injection Program at Texas A&M University for in vitro blastocyst production during the past 3 years.
October 8, 2016: Veterinary Clinics of North America. Equine Practice
Vladimir Isachenko, Karl Sterzik, Robert Maettner, Evgenia Isachenko, Plamen Todorov, Gohar Rahimi, Peter Mallmann, Erwin Strehler, Igor Pereligin, José Luis Alabart, Markus Merzenich
In natural conditions the oocyte and embryo are subjected to ever changing dynamic processes. However, the routine assisted reproductive technologies today involve the use of static in vitro culture systems. Objective was to determine whether there is any difference in the viability of embryos after in vitro culture under static and mechanical micro-vibration condition. It was evaluated the viability of embryonic cells (9,624 embryos) generated from 4,436 couples after in vitro culture. For Groups <29 years, 30-34 years, 35-39 years and >40 years, the following rate of high quality embryos without fragmentation (2 to 4 blastomeres on Day 2; 6 to 8 blastomeres and compacting morula on Day 3; blastocyst, expanded and hatching blastocyst on Day 5) was detected (static vs...
October 7, 2016: Cell Transplantation
Robert S Illingworth, Jurriaan J Hölzenspies, Fabian V Roske, Wendy A Bickmore, Joshua M Brickman
Mouse embryonic stem cells (ESCs), like the blastocyst from which they are derived, contain precursors of the epiblast (Epi) and primitive endoderm (PrEn) lineages. While transient in vivo, these precursor populations readily interconvert in vitro. We show that altered transcription is the driver of these coordinated changes, known as lineage priming, in a process that exploits novel polycomb activities. We find that intragenic levels of the polycomb mark H3K27me3 anti-correlate with changes in transcription, irrespective of the gene's developmental trajectory or identity as a polycomb target...
October 10, 2016: ELife
Natalie K Binder, Jemma Evans, Lois A Salamonsen, David K Gardner, Tu'uhevaha J Kaitu'u-Lino, Natalie J Hannan
Embryo implantation requires synchronized dialogue between the receptive endometrium and activated blastocyst via locally produced soluble mediators. During the mid-secretory (MS) phase of the menstrual cycle, increased glandular secretion into the uterine lumen provides important mediators that modulate the endometrium and support the conceptus during implantation. Previously we demonstrated the importance of vascular endothelial growth factor (VEGF) in the human uterus, particularly with respect to embryo implantation...
2016: PloS One
Nilay Kuşcu, Mariano Bizzarri, Arturo Bevilacqua
Myo-inositol (myo-Ins) has a physiological role in mammalian gametogenesis and embryonic development and a positive clinical impact on human medically assisted reproduction. We have previously shown that mouse embryo exposure to myo-Ins through preimplantation development in vitro increases proliferation activity and blastocyst production, representing an improvement in culture conditions. We have herein investigated biochemical mechanisms elicited by myo-Ins in preimplantation embryos and evaluated myo-Ins effects on postimplantation/postnatal development...
2016: International Journal of Endocrinology
Isao Matsuo, Ryuji Hiramatsu
In most mammals, embryonic development and growth proceed in the maternal uterus. Mouse late blastocyst embryos implant on the uterine epithelium around embryonic day (E)4.5, and immediately afterward the whole embryo's shape is dynamically changed from a bowl-like shape to an elongated egg-cylinder until E5.5. Concurrently, mouse anterior-posterior (A-P) axis polarization occurs by the emergence of distal visceral endoderm (DVE) cells at the cellular and molecular levels as the proximal-distal (P-D) axis. The embryonic growth and axis polarization are considered to be controlled primarily by multiple growth factors' signaling...
September 30, 2016: Mechanisms of Development
A Mishra, I J Reddy, Psp Gupta, S Mondal
The objective of this study was to find out the impact of L-carnitine (10 mM) on developmental regulation of preimplantation sheep embryos cultured in vitro when supplemented in maturation medium and post-fertilization medium separately. Subsequent objective was to observe the L-carnitine-mediated alteration in expression of apoptotic genes (Bcl2, Bax, Casp3 and PCNA) in sheep oocytes and developing embryos produced in vitro. Oocytes matured with L-carnitine showed significantly (p < .05) higher cleavage (67...
September 30, 2016: Reproduction in Domestic Animals, Zuchthygiene
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