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Blastocyst culture

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https://www.readbyqxmd.com/read/28093041/blastocyst-development-in-a-single-medium-compared-to-sequential-media
#1
Ioannis A Sfontouris, Efstratios M Kolibianakis, George T Lainas, George K Petsas, Basil C Tarlatzis, Trifon G Lainas
The aim of the present study was to compare blastocyst formation rates after embryo culture in a single medium (Global) as compared to sequential media (ISM1/BlastAssist). In this prospective trial with sibling oocytes, 542 metaphase II (MII) oocytes from 31 women were randomly and equally divided to be fertilized and cultured to the blastocyst stage in either sequential media (ISM1/BlastAssist; n = 271 MII oocytes) or a single medium (Global; n = 271 MII oocytes). In both groups, embryos were cultured in an interrupted fashion with media changes on day 3...
January 1, 2017: Reproductive Sciences
https://www.readbyqxmd.com/read/28089671/pdgfr%C3%AE-cells-in-embryonic-stem-cell-cultures-represent-the-in%C3%A2-vitro-equivalent-of-the-pre-implantation-primitive-endoderm-precursors
#2
Antonio Lo Nigro, Anchel de Jaime-Soguero, Rita Khoueiry, Dong Seong Cho, Giorgia Maria Ferlazzo, Ilaria Perini, Vanesa Abon Escalona, Xabier Lopez Aranguren, Susana M Chuva de Sousa Lopes, Kian Peng Koh, Pier Giulio Conaldi, Wei-Shou Hu, An Zwijsen, Frederic Lluis, Catherine M Verfaillie
In early mouse pre-implantation development, primitive endoderm (PrE) precursors are platelet-derived growth factor receptor alpha (PDGFRα) positive. Here, we demonstrated that cultured mouse embryonic stem cells (mESCs) express PDGFRα heterogeneously, fluctuating between a PDGFRα+ (PrE-primed) and a platelet endothelial cell adhesion molecule 1 (PECAM1)-positive state (epiblast-primed). The two surface markers can be co-detected on a third subpopulation, expressing epiblast and PrE determinants (double-positive)...
January 5, 2017: Stem Cell Reports
https://www.readbyqxmd.com/read/28073983/a-novel-embryo-culture-media-supplement-that-improves-pregnancy-rates-in-mice
#3
A R Highet, T Bianco-Miotto, K G Pringle, A Peura, S Bent, J Zhang, M B Nottle, J G Thompson, C T Roberts
The preimplantation embryo in vivo is exposed to numerous growth factors in the female reproductive tract, which are not recapitulated in embryo culture media in vitro The IGF2 and plasminogen activator systems facilitate blastocyst development. We hypothesized that the addition of IGF2 in combination with urokinase plasminogen activator (uPA) and plasminogen could improve rates of blastocyst hatching and implantation in mice. B6BcF1 and CBAB6F2 mouse embryos were divided into one of four supplemented culture media treatment groups: (1) control (media only); (2) 12...
March 2017: Reproduction: the Official Journal of the Society for the Study of Fertility
https://www.readbyqxmd.com/read/28069902/wnt-regulation-of-embryonic-development-likely-involves-pathways-independent-of-nuclear-ctnnb1
#4
Paula Tribulo, James I Moss, Manabu Ozawa, Zongliang Jiang, Cindy Tian, Peter J Hansen
The bovine was used to examine potential for WNT signaling to affect the preimplantation embryo. Expression of seven key genes involved in canonical WNT signaling declined to a nadir at the morula or blastocyst stage. Expression of 80 genes associated with WNT signaling in the morula and inner cell mass (ICM) and trophectoderm (TE) of the blastocyst was also evaluated. Many genes associated with WNT signaling were characterized by low transcript abundance. Seven genes were different between ICM and TE, and all of them were overexpressed in TE as compared to ICM, including WNT6, FZD1, FZD7, LRP6, PORCN, APC, and SFRP1...
January 9, 2017: Reproduction: the Official Journal of the Society for the Study of Fertility
https://www.readbyqxmd.com/read/28064422/improved-detection-of-mineral-oil-toxicity-using-an-extended-mouse-embryo-assay
#5
Alessandra J Ainsworth, Jolene R Fredrickson, Dean E Morbeck
PURPOSE: Successful in vitro fertilization (IVF) relies on sound laboratory methods and culture conditions which depend on sensitive quality control (QC) testing. This study aimed to improve the sensitivity of mouse embryo assays (MEA) for detection of mineral oil toxicity. METHODS: Five experiments were conducted to study modifications of the standard mouse embryo assay (MEA) in order to improve sensitivity using clinical grade mineral oil with known peroxide concentrations...
January 7, 2017: Journal of Assisted Reproduction and Genetics
https://www.readbyqxmd.com/read/28057448/improved-cryotolerance-and-developmental-competence-of-human-oocytes-matured-in-vitro-by-transient-hydrostatic-pressure-treatment-prior-to-vitrification
#6
Ruihuan Gu, Yun Feng, Song Guo, Shen Zhao, Xiaowei Lu, Jing Fu, Xiaoxi Sun, Yijuan Sun
BACKGROUND: At present, the metaphase II (MII) oocytes have a very special structure that leads to complex difficulties associated with its vitrification, and their efficacy still needs a large amount of study to observe. The present study was to investigate whether transient hydrostatic pressure (THP), which was utilized for oocytes before vitrification, had positive effect on the oocytes' developmental ability and reactive oxygen species, and had no damage on meiotic spindle, zona pellucida, and DNA copy number...
January 2, 2017: Cryobiology
https://www.readbyqxmd.com/read/28050958/non-invasive-prediction-of-blastocyst-implantation-ongoing-pregnancy-and-live-birth-by-mass-spectrometry-lipid-fingerprinting
#7
Edson Borges, Daniela P A F Braga, Amanda Souza Setti, Daniela A Montanni, Elaine Cristina Cabral, Marcos N Eberlin, Edson G Lo Turco, Assumpto Iaconelli
OBJECTIVE: To identify lipid markers of blastocyst implantation and ongoing pregnancy by day three culture medium mass spectrometry (MS) fingerprinting. METHODS: For this study, 33 culture media samples were harvested on day three, from 22 patients undergoing day five embryo transfers. All embryos achieved the blastocyst stage and were split into groups based on their implantation (Negative Implantation, n= 14 and Positive Implantation, n= 19). The positive implantation cycles resulted in successful ongoing pregnancies...
December 1, 2016: JBRA Assisted Reproduction
https://www.readbyqxmd.com/read/28050953/mir-142-3p-as-a-biomarker-of-blastocyst-implantation-failure-a-pilot-study
#8
Edson Borges, Amanda Souza Setti, Daniela P A F Braga, Murilo V Geraldo, Rita de Cássia S Figueira, Assumpto Iaconelli
OBJECTIVE: This study aims to find whether microRNAs (miRNAs) detected in the culture medium of embryos produced in vitro could be potential biomarkers of embryo implantation. METHODS: Culture media samples from 36 embryos, derived from patients undergoing intracytoplasmic sperm injection (ICSI) in a private university-affiliated IVF center, were collected between January/2015 and November/2015. Samples were collected on day three and embryo transfers were performed on day five and all embryos reached the blastocyst stage...
December 1, 2016: JBRA Assisted Reproduction
https://www.readbyqxmd.com/read/28049782/derivation-and-culture-of-extra-embryonic-endoderm-stem-cell-lines
#9
Peter Rugg-Gunn
Whereas embryonic stem (ES) cells are isolated from the embryonic lineage of the blastocyst, other stable stem cell lines can be derived from the extraembryonic tissues of the early mouse embryo. Trophoblast stem (TS) cells are derived from trophectoderm and early postimplantation trophoblast, and extraembryonic endoderm stem (XEN) cells are derived from primitive endoderm. The derivation of XEN cell lines from 3.5-dpc mouse blastocysts, described here, is similar to the derivation of TS cell lines. TS and XEN cells can self-renew in vitro and differentiate in vitro and in chimeras (in vivo) in a lineage-appropriate manner, showing the developmental potential of their origin, thus providing important models to study the mouse extraembryonic lineages...
January 3, 2017: Cold Spring Harbor Protocols
https://www.readbyqxmd.com/read/28043370/assessment-of-the-effect-of-adding-l-carnitine-and-or-resveratrol-to-maturation-medium-before-vitrification-on-in%C3%A2-vitro-matured-calf-oocytes
#10
José Felipe Sprícigo, Roser Morató, Núria Arcarons, Marc Yeste, Margot Alves Dode, Manuel López-Bejar, Teresa Mogas
Cryopreservation may lead bovine oocytes to undergo morphological changes and functional damage due to the high-lipid content in the cytoplasm and the formation of reactive oxygen species. Against this background, the present study aimed to improve the cryotolerance and developmental competence of prepubertal calf oocytes by adding L-carnitine (LC) and/or resveratrol (R) to the IVM medium, as the former is involved in lipid metabolism and both are able to scavenge reactive oxygen species. With this purpose, different quality and functional oocyte parameters, such as spindle and chromosome configuration, DNA integrity, caspase activity, and the profile of genes involved in lipid metabolism and oxidative stress were evaluated in IVM bovine oocytes before or after vitrification/warming...
February 2017: Theriogenology
https://www.readbyqxmd.com/read/28043361/time-of-early-cleavage-affects-the-developmental-potential-of-feline-preimplantation-embryos-in%C3%A2-vitro
#11
Małgorzata Ochota, Wojciech Niżański
The study compared developmental competence of embryos, based on the timing of the first cleavage and morula formation, with their subsequent ability to reach the blastocyst stage and the resulting blastocyst morphology and quality. Cleaved embryos were separated at 18 hours post insemination (hpi), 24 hpi, and 30 hpi and then cultured in droplets to follow the individual embryo development. The significantly higher percentage of the blastocyst formation was noted in the group I of embryos cleaving within less than 18 hpi (12...
February 2017: Theriogenology
https://www.readbyqxmd.com/read/28042416/which-stage-of-mouse-embryos-is-more-appropriate-for-vitrification
#12
Nasibeh Ghandy, Abbas Ali Karimpur Malekshah
BACKGROUND: Vitrification has been shown as one of the most effective methods of cryopreservation for mammalian embryos. However, there is no consensus which stage of embryonic development is the most appropriate for vitrification with subsequent maximal development after thawing. This study was carried out to explore and compare the effect(s) of vitrification on mouse 2-cell, 4-cell, 8-cell, morula and blastocyst stage embryos and subsequent blast formation and hatching after thawing...
January 2017: International Journal of Fertility & Sterility
https://www.readbyqxmd.com/read/28007046/maternal-effect-gene-expression-in-porcine-metaphase-ii-oocytes-and-embryos-in-vitro-effect-of-epidermal-growth-factor-interleukin-1%C3%AE-and-leukemia-inhibitory-factor
#13
Marta Wasielak, Teresa Więsak, Iwona Bogacka, Beenu Moza Jalali, Marek Bogacki
Maternal effect genes (MEG) play a crucial role in early embryogenesis. In vitro culture conditions may affect MEG expression in porcine oocytes and embryos. We investigated whether in vitro culture medium supplementation with epidermal growth factor (EGF), IL-1β or LIF (leukemia inhibitory factor) affects the mRNA level of ZAR-1 (zygote arrest 1), NPM2 (nucleoplasmin 2) and DPPA3 (developmental associated protein 3) in porcine MII oocytes and embryos. Cumulus-oocyte complexes (COCs) were matured in NCSU-37 medium (control) or in NCSU-37 with EGF 10 ng/ml, IL-1β 10 ng/ml or LIF 50 ng/ml...
December 23, 2016: Zygote: the Biology of Gametes and Early Embryos
https://www.readbyqxmd.com/read/28004977/establishment-of-transgenic-porcine-fibroblasts-expressing-a-human-klotho-gene-and-its-effects-on-gene-expression-and-preimplantation-development-of-cloned-embryos
#14
Sanghoon Lee, Joon Ho Moon, Kilyoung Song, Anukul Taweechaipaisankul, Young Kwang Jo, Hyun Ju Oh, Se Chang Park, Byeong Chun Lee
Even though the functions of the klotho gene in aging of small animals such as mice have been well investigated, studies using large animal models such as pigs, which have many similarities to humans, have been limited due to the absence of cell lines or animal models. Therefore, the objective of this study was to generate porcine cell lines overexpressing human klotho (hKlotho) and tetracycline (Tet)-inducible hKlotho and to produce cloned embryos from these cell lines. We designed vectors for hKlotho overexpression (CA-Klotho) under control of CMV enhancer/chicken β-actin (CAG) promoter and Tet-inducible hKlotho overexpression (Tet-Klotho, under control of doxycycline-dependent promoter)...
January 2017: DNA and Cell Biology
https://www.readbyqxmd.com/read/27989770/human-naive-pluripotent-stem-cells-model-x-chromosome-dampening-and-x-inactivation
#15
Anna Sahakyan, Rachel Kim, Constantinos Chronis, Shan Sabri, Giancarlo Bonora, Thorold W Theunissen, Edward Kuoy, Justin Langerman, Amander T Clark, Rudolf Jaenisch, Kathrin Plath
Naive human embryonic stem cells (hESCs) can be derived from primed hESCs or directly from blastocysts, but their X chromosome state has remained unresolved. Here, we show that the inactive X chromosome (Xi) of primed hESCs was reactivated in naive culture conditions. Like cells of the blastocyst, the resulting naive cells contained two active X chromosomes with XIST expression and chromosome-wide transcriptional dampening and initiated XIST-mediated X inactivation upon differentiation. Both establishment of and exit from the naive state (differentiation) happened via an XIST-negative XaXa intermediate...
January 5, 2017: Cell Stem Cell
https://www.readbyqxmd.com/read/27989758/low-level-of-mono-2-ethylhexyl-phthalate-reduces-oocyte-developmental-competence-in-association-with-impaired-gene-expression
#16
D Kalo, Z Roth
Di-(2-ethylhexyl) phthalate (DEHP) and its metabolite, mono-(2-ethylhexyl) phthalate (MEHP), are reproductive toxicants. However, disruptive effects of MEHP at low concentrations on the oocyte and developing blastocyst are unknown. Previously, we detected low levels of MEHP in follicular fluid aspirated from DEHP-treated cows associated with reduced estradiol levels. Moreover, the MEHP concentrations found were similar to those reported for follicular fluid aspirated from women who have undergone IVF cycles...
December 15, 2016: Toxicology
https://www.readbyqxmd.com/read/27980237/prediction-of-blastocyst-development-and-implantation-potential-in-utero-based-on-the-third-cleavage-and-compaction-times-in-mouse-pre-implantation-embryos
#17
Jihyun Kim, Seok Hyun Kim, Jin Hyun Jun
Cytokinesis and cell division during pre-implantation embryonic development occur as an orchestrated spatiotemporal program. Cleavage, compaction, and blastulation in pre-implantation embryos are essential for successful implantation and pregnancy. Their alteration is associated with chromosomal imbalance and loss of developmental competence. In this study, we evaluated the time of cleavage and compaction as predictors for in vitro pre- and peri-implantation development and in utero implantation potential by time-lapse monitoring...
December 16, 2016: Journal of Reproduction and Development
https://www.readbyqxmd.com/read/27955719/promotion-of-glucose-utilization-by-insulin-enhances-granulosa-cell-proliferation-and-developmental-competence-of-porcine-oocyte-grown-in-vitro
#18
Nobuhiko Itami, Yasuhisa Munakata, Koumei Shirasuna, Takehito Kuwayama, Hisataka Iwata
In vitro culture of the oocyte granulosa cell complexes (OGCs) from early antral follicles (EAFs) shows granulosa cell (GC) proliferation, but to a lesser extent than that observed in vivo during follicle development. As the number of GCs closely relates to energy sufficiency of the oocytes, enhancement of GC proliferation influences oocyte development. GC proliferation depends on glycolysis and insulin-mediated AKT/mTOR signaling pathway; therefore, addition of culture medium containing insulin and glucose may potentially promote GC proliferation and hence improve oocyte development...
February 2017: Zygote: the Biology of Gametes and Early Embryos
https://www.readbyqxmd.com/read/27943152/oocyte-maturation-embryo-development-and-gene-expression-following-two-different-methods-of-bovine-cumulus-oocyte-complexes-vitrification
#19
Mehdi Azari, Mojtaba Kafi, Bita Ebrahimi, Roya Fatehi, Mahboobeh Jamalzadeh
OBJECTIVE: To examine the maturational competence, embryo development and expression of genes involved in oocyte maturation and cumulus expansion (GDF9, BMP15, HAS2, TNFAIP6, FGF17 and FSHr) following two standard methods of bovine COCs vitrification. METHODS: Bovine cumulus-oocyte complexes (COCs) were aspirated from slaughtered ovaries and then distributed into three groups: non-vitrified COCs (control), vitrification 1 group (V1); vitrification was performed by 15% ethylene glycol (EG) and 15% DMSO in holding media (TCM-199 with 20% FCS); and vitrification 2 group (V2); vitrification was performed by 40% EG in holding media...
December 10, 2016: Veterinary Research Communications
https://www.readbyqxmd.com/read/27943109/morphokinetics-of-vitrified-and-warmed-blastocysts-predicts-implantation-potential
#20
T Ebner, P Oppelt, E Radler, C Allerstorfer, A Habelsberger, R B Mayer, O Shebl
PURPOSE: It was studied whether morphokinetics of blastocoele re-expansion and hatching in vitrified-warmed blastocysts is predictive of implantation, clinical pregnancy, and live birth. METHODS: In 144 patients aiming for single warmed blastocyst transfer, blastocysts were cultured in a new time-lapse system (Miri® TL) immediately after warming. Video sequences with an image interval of 5 min were annotated and the corresponding morphokinetic variables were correlated with pregnancy outcome...
December 9, 2016: Journal of Assisted Reproduction and Genetics
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