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https://www.readbyqxmd.com/read/29175537/vitrification-of-human-pronuclear-oocytes-by-direct-plunging-into-cooling-agent-non-sterile-liquid-nitrogen-vs-sterile-liquid-air
#1
Vladimir Isachenko, Plamen Todorov, Akerke Seisenbayeva, Yerzhan Toishibekov, Evgenia Isachenko, Gohar Rahimi, Peter Mallmann, Dolores Foth, Markus Merzenich
In fact, a full sterilization of commercially-produced liquid nitrogen contaminated with different pathogens is not possible. The aim of this study was to compare the viability of human pronuclear oocytes subjected to cooling by direct submerging of open carrier in liquid nitrogen versus submerging in clean liquid air (aseptic system). One- and three-pronuclei stage embryos (n = 444) were cryopreserved by direct plunging into liquid nitrogen (vitrified) in ethylene glycol (15%), dimethylsulphoxide (15%) and 0...
November 22, 2017: Cryobiology
https://www.readbyqxmd.com/read/29149327/focused-time-lapse-analysis-reveals-novel-aspects-of-human-fertilization-and-suggests-new-parameters-of-embryo-viability
#2
G Coticchio, M Mignini Renzini, P V Novara, M Lain, E De Ponti, D Turchi, R Fadini, M Dal Canto
STUDY QUESTION: Can focused application of time-lapse microscopy (TLM) lead to a more detailed map of the morphokinetics of human fertilization, revealing novel or neglected aspects of this process? SUMMARY ANSWER: Intensive harnessing of TLM reveals novel or previously poorly characterised phenomena of fertilization, such as a cytoplasmic wave (CW) preceding pronuclear formation and kinetics of pronuclear chromatin polarization, thereby suggesting novel non-invasive biomarkers of embryo quality...
November 15, 2017: Human Reproduction
https://www.readbyqxmd.com/read/29118980/sirt1-dependent-modulation-of-methylation-and-acetylation-of-histone-h3-on-lysine-9-h3k9-in-the-zygotic-pronuclei-improves-porcine-embryo-development
#3
Katerina Adamkova, Young-Joo Yi, Jaroslav Petr, Tereza Zalmanova, Kristyna Hoskova, Pavla Jelinkova, Jiri Moravec, Milena Kralickova, Miriam Sutovsky, Peter Sutovsky, Jan Nevoral
Background: The histone code is an established epigenetic regulator of early embryonic development in mammals. The lysine residue K9 of histone H3 (H3K9) is a prime target of SIRT1, a member of NAD(+)-dependent histone deacetylase family of enzymes targeting both histone and non-histone substrates. At present, little is known about SIRT1-modulation of H3K9 in zygotic pronuclei and its association with the success of preimplantation embryo development. Therefore, we evaluated the effect of SIRT1 activity on H3K9 methylation and acetylation in porcine zygotes and the significance of H3K9 modifications for early embryonic development...
2017: Journal of Animal Science and Biotechnology
https://www.readbyqxmd.com/read/29074360/perivitelline-threads-in-cleavage-stage-human-embryos-observations-using-time-lapse-imaging
#4
Louise Kellam, Laura M Pastorelli, Angel M Bastida, Amy Senkbeil, Sue Montgomery, Simon Fishel, Alison Campbell
Time-lapse imaging of the human preimplantation embryo in vitro has revealed a transient phenomenon involving the appearance of perivitelline threads, commonly observed at the two-cell stage. These threads span the perivitelline space, arising at the specific area where the cytoplasmic membrane contacts the zona pellucida, before any perivitelline space is formed. The threads persist as the cytoplasmic membrane retracts from the zona pellucida to form the first cleavage furrow. In this observational report, these structures and their incidence are described...
September 28, 2017: Reproductive Biomedicine Online
https://www.readbyqxmd.com/read/29034763/pronuclear-pattern-does-not-predict-morphokinetics-behavior-in-human-embryos
#5
Azita Faramarzi, Mohammad Ali Khalili, Marjan Omidi, Azam Agha-Rahimi, Fatemeh Taheri
The purpose was to investigate the correlation between pronuclei (PN) morphology and morphokinetic behaviors of derived embryos with time lapse monitoring (TLM) in assisted reproduction setting. Over time, PN morphology from PN appearance (PNA) to PN fading (PNF), PNF according to size, contact, number and position of nuclear precursor bodies (NPBs) within each PN and morphokinetics variables, including absolute time points, relative timing parameters, cleavage patterns and arrest rate, were evaluated using TLM...
October 16, 2017: Gynecological Endocrinology
https://www.readbyqxmd.com/read/29034232/maternally-contributed-folate-receptor-1-is-expressed-in-ovarian-follicles-and-contributes-to-preimplantation-development
#6
Trine Strandgaard, Solveig Foder, Anders Heuck, Erik Ernst, Morten S Nielsen, Karin Lykke-Hartmann
Folates have been shown to play a crucial role for proper development of the embryo as folate deficiency has been associated with reduced developmental capacity such as increased risk of fetal neural tube defects and spontanous abortion. Transcripts encoding the reduced folate carrier RFC1 (SLC19A1 protein) and the high-affinity folate receptor FOLR1 are expressed in oocytes and preimplantation embryos, respectively. In this study, we observed maternally contributed FOLR1 protein during mouse and human ovarian follicle development, and 2-cell mouse embryos...
2017: Frontiers in Cell and Developmental Biology
https://www.readbyqxmd.com/read/28955073/comparison-of-embryo-morphokinetics-following-intracytoplasmic-sperm-injection-in-smoker-and-non-smoker-couples-are-the-results-different
#7
Ahmet Salvarci, Ali Sami Gurbuz, Sukru Uzman, Melek Kaya, Huseyin Gorkemli
OBJECTIVE: To assess early embryo development via time-lapse in smokers and non-smokers. METHODS: The retrospective study was conducted at Novafertil IVF centers in Konya, Turkey and comprised oocytes of both smoker and non-smoker couples subjected to in vitro fertilisation / introcytoplasmic sperm injection from 2012 to 2015. Age, basal follicle-stimulating hormone, number of stimulation days, amount of gonadotropin used, number of metaphase II oocytes, number of embryos transferred and pregnancy, abortus and clinical pregnancy rates were noted...
October 2017: JPMA. the Journal of the Pakistan Medical Association
https://www.readbyqxmd.com/read/28886319/morphokinetic-analysis-of-human-embryo-development-and-its-relationship-to-the-female-age-a-retrospective-time-lapse-imaging-study
#8
N Akhter, M Shahab
Time-lapse imaging technique has provided embryologists with a unique chance of studying the embryo morphokineticsto select the most viable embryos for implantation in the field of IVF (In vitro fertilization).The conventional method of morphological evaluations has proved that maternal age affects the human embryo quality. This retrospective study carried out at Islamabad Clinic Serving Infertile Couples, Islamabad, Pakistan mainly focuses on the effect of female age on human embryo morphokinetics. A total number of 200 patients undergoing ICSI treatment cycles at the clinic were selected for the study and divided into five age groups (< 26, 26-30, 31-35, 36-40, and > 40 years)...
August 30, 2017: Cellular and Molecular Biology
https://www.readbyqxmd.com/read/28855524/rnai-combining-sleeping-beauty-transposon-system-inhibits-ex-vivo-expression-of-foot-and-mouth-disease-virus-vp1-in-transgenic-sheep-cells
#9
Shoulong Deng, Guangdong Li, Kun Yu, Xiuzhi Tian, Feng Wang, Wenting Li, Wuqi Jiang, Pengyun Ji, Hongbing Han, Juncai Fu, Xiaosheng Zhang, Jinlong Zhang, Yixun Liu, Zhengxing Lian, Guoshi Liu
Foot and mouth disease, which is induced by the foot and mouth disease virus (FMDV), takes its toll on the cloven-hoofed domestic animals. The VP1 gene in FMDV genome encodes the viral capsid, a vital element for FMDV replication. Sleeping Beauty (SB) is an active DNA-transposon system responsible for genetic transformation and insertional mutagenesis in vertebrates. In this study, a conserved VP1-shRNA which specifically targets the ovine FMDV-VP1 gene was constructed and combined with SB transposase and transposon...
August 30, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28809019/lentiviral-mediated-transgenesis-in-songbirds
#10
Wan-Chun Liu, Marian Hruska-Plochan, Atsushi Miyanohara
Transgenesis involves the insertion of an exogenous gene into an animal's genome, which allows the identification of the expressed phenotypes in brain function or behavior. Lentiviral-mediated transgenesis offers unique transduction potency making it possible to deliver and stably integrate transgenes into a wide variety of dividing and nondividing cells. The ability to establish long-term expression of such transgenes allows their use for transgenesis which is especially useful in organisms lacking quality pluripotent stem cell lines and which is otherwise difficult to produce via traditional pronuclear microinjection, such as songbirds...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28795048/evaluation-of-human-embryo-development-in-in-vitro-fertilization-and-intracytoplasmic-sperm-injection-fertilized-oocytes-a-time-lapse-study
#11
Hyung Jun Kim, Hye Jin Yoon, Jung Mi Jang, Won Don Lee, San Hyun Yoon, Jin Ho Lim
OBJECTIVE: We investigated whether the insemination method (in vitro fertilization [IVF] or intracytoplasmic sperm injection [ICSI]) affected morphokinetic events and abnormal cleavage events in embryonic development. METHODS: A total of 1,830 normal fertilized embryos were obtained from 272 IVF and ICSI cycles that underwent ovum retrieval culture using a time-lapse system (Embryoscope) from June 2013 to March 2015. All embryos were investigated by a detailed time-lapse analysis that measured the developmental events in the hours after IVF or ICSI insemination...
June 2017: Clinical and Experimental Reproductive Medicine
https://www.readbyqxmd.com/read/28765300/pronuclear-transplantation-in-the-mouse-embryo
#12
James McGrath, Davor Solter, Jeff Mann
This protocol describes an example of complete zygote enucleation and transplantation of male and female pronuclei; however, single pronuclei can also be removed and transplanted. In this method, pronuclei are removed without penetrating the plasma membrane of the zygote. Instead, they are withdrawn individually or together into a membrane-bound karyoplast that can then be fused with a recipient enucleated zygote using inactivated Sendai virus or electrofusion. Preincubation of the embryos in the presence of the cytoskeletal inhibitors cytochalasin B and colcemid is critical for the survival of the embryos during this microsurgical procedure...
August 1, 2017: Cold Spring Harbor Protocols
https://www.readbyqxmd.com/read/28757075/dna-demethylation-pattern-of-in-vitro-fertilized-and-cloned-porcine-pronuclear-stage-embryos
#13
Xiaowei Nie, Qiang Liu, Ronggen Wang, Wenjie Sheng, Xiaokang Li, Manling Zhang, Yong Jin, Lihua Zhao, Daorong Hou, Ning Yang, Zhaoqiang Wu, Yifan Dai, Rongfeng Li
Recent studies in mice showed that the Ten-eleven translocation Enzymes (TET) family is involved in the active DNA demethylation. The isotype TET-3 is responsible for the conversion of 5mc (5-methylcytosine) to 5hmc (5-hydroxymethylcytosine) at the pronuclear stages of mouse embryo. This study was performed to investigate the pattern of methylation change and the role of TET family in the demethylation process of porcine in-vitro fertilization (IVF) and somatic cell nuclear transfer (SCNT) derived embryo. Bisulfite-sequencing PCR (BSP) and DNA glucosylation and digestion before quantitative PCR (qGluMS-PCR) were done to evaluate the exact change of methylation during porcine pronuclear stages...
October 2017: Clinica Chimica Acta; International Journal of Clinical Chemistry
https://www.readbyqxmd.com/read/28643254/gene-targeting-in-rabbits-single-step-generation-of-knock-out-rabbits-by-microinjection-of-crispr-cas9-plasmids
#14
Yoshihiro Kawano, Arata Honda
The development of genome editing technology has allowed gene disruptions to be achieved in various animal species and has been beneficial to many mammals. Gene disruption using pluripotent stem cells is difficult to achieve in rabbits, but thanks to advances in genome editing technology, a number of gene disruptions have been conducted. This paper describes a simple and easy method for carrying out gene disruptions in rabbits using CRISPR/Cas9 in which the gene to be disrupted is marked, the presence or absence of off-target candidates is checked, and a plasmid allowing simultaneous expression of Cas9 and sgRNA is constructed...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28643251/genome-editing-in-mouse-and-rat-by-electroporation
#15
Takehito Kaneko
Many knock-out/knock-in mouse and rat strains have been produced by genome editing techniques using engineered endonucleases, including zinc finger nuclease (ZFN), transcription activator-like effector nuclease (TALEN), or clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9. Microinjection of engineered endonucleases into pronuclear-stage embryos is required to produce genome-edited rodents and the development of easy, rapid, and high-efficiency methods that do not require special skills such as microinjection is needed...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28601410/total-number-of-oocytes-and-zygotes-are-predictive-of-live-birth-pregnancy-in-fresh-donor-oocyte-in%C3%A2-vitro-fertilization-cycles
#16
Eduardo Hariton, Keewan Kim, Sunni L Mumford, Marissa Palmor, Pietro Bortoletto, Eden R Cardozo, Anatte E Karmon, Mary E Sabatini, Aaron K Styer
OBJECTIVE: To evaluate the association of oocyte donor-recipient characteristics, oocyte donor response, and live birth pregnancy rate following fresh donor oocyte IVF-ET. DESIGN: Retrospective cohort study. SETTING: Academic reproductive medicine practice. PATIENT(S): Two hundred thirty-seven consecutive fresh donor oocyte IVF-ET cycles from January 1, 2007 to December 31, 2013 at the Massachusetts General Hospital Fertility Center...
August 2017: Fertility and Sterility
https://www.readbyqxmd.com/read/28545999/a-comparative-analysis-of-the-efficacy-of-three-cryopreservation-protocols-on-the-survival-of-in%C3%A2-vitro-derived-cattle-embryos-at-pronuclear-and-blastocyst-stages
#17
Van Huong Do, Simon Walton, Sally Catt, Andrew W Taylor-Robinson
The effectiveness of three cryopreservation protocols (slow freezing, short equilibration vitrification and long equilibration vitrification) on in vitro-derived cattle embryos at expanded blastocyst and pronuclear stages was compared. 199 expanded blastocysts of good quality were assigned randomly into four treatment groups [control, non-cryopreserved (fresh, unfrozen); and the three cryopreservation methods]. The re-expansion of the cryopreserved blastocysts after 24 h in vitro culture was similar to that of the fresh control group...
May 22, 2017: Cryobiology
https://www.readbyqxmd.com/read/28475125/melatonin-promotes-the-in-vitro-development-of-microinjected-pronuclear-mouse-embryos-via-its-anti-oxidative-and-anti-apoptotic-effects
#18
Xiuzhi Tian, Feng Wang, Lu Zhang, Pengyun Ji, Jing Wang, Dongying Lv, Guangdong Li, Menglong Chai, Zhengxing Lian, Guoshi Liu
CRISPR/Cas9 (Clustered regularly interspaced short palindromic repeats) combined with pronuclear microinjection has become the most effective method for producing transgenic animals. However, the relatively low embryo developmental rate limits its application. In the current study, it was observed that 10(-7) M melatonin is considered an optimum concentration and significantly promoted the in vitro development of murine microinjected pronuclear embryos, as indicated by the increased blastocyst rate, hatching blastocyst rate and blastocyst cell number...
May 5, 2017: International Journal of Molecular Sciences
https://www.readbyqxmd.com/read/28456971/use-of-histone-k-m-mutants-for-the-analysis-of-transcriptional-regulation-in-mouse-zygotes
#19
Keisuke Aoshima, Takashi Kimura, Yuki Okada
Histone modifications are dramatically altered during the pronuclear (PN) stage of zygotes, and more markedly in paternal than maternal pronuclei. Among various types of histone modifications, lysine methylation exhibits the most dynamic changes in the PN stage . To analyze the physiological functions of histone methylations, it is therefore important to elucidate the mechanism of epigenetic reprogramming. However, loss-of-function approaches using mutant histones whose lysine residues have been substituted with arginine residues are unable to erase histone modifications at all levels, since they are incapable of entirely replacing endogenous histones...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28448636/successful-vitrification-of-pronuclear-stage-pig-embryos-with-a-novel-cryoprotective-agent-carboxylated-%C3%AE%C2%B5-poly-l-lysine
#20
Maki Kamoshita, Tsubasa Kato, Katsuyoshi Fujiwara, Takafumi Namiki, Kazuaki Matsumura, Suong-Hyu Hyon, Junya Ito, Naomi Kashiwazaki
Vitrification is a powerful tool for the efficient production of offspring derived from cryopreserved oocytes or embryos in mammalian species including domestic animals. Genome editing technologies such as transcription activator-like effector nucleases (TALENs) and clustered regularly interspaced short palindromic repeats (CRISPR)/ CRISPR-associated (Cas)9 are now available even for domestic species, suggesting that the vitrification of embryos at the pronuclear stage (PN) will be more important because they could provide genomic host cells to be targeted by TALENs or CRISPR/Cas9...
2017: PloS One
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