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Single step embryo culture

Séverine Le Gac, Verena Nordhoff
The optimization of in-vitro culture conditions and the selection of the embryo(s) with the highest developmental competence are essential components in an ART program. Culture conditions are manifold and they underlie not always evidence-based research but also trends entering the IVF laboratory. At the moment, the idea of using sequential media according to the embryo requirements has been given up in favor of the use of single step media in an uninterrupted manner due to practical issues such as time-lapse incubators...
September 27, 2016: Molecular Human Reproduction
Ioannis A Sfontouris, Wellington P Martins, Carolina O Nastri, Iara G R Viana, Paula A Navarro, Nick Raine-Fenning, Sheryl van der Poel, Laura Rienzi, Catherine Racowsky
PURPOSE: The purpose of this study was to undertake a review of the available evidence comparing the use of a single medium versus sequential media for embryo culture to the blastocyst stage in clinical IVF. METHODS: We searched the Cochrane Central, PubMed, Scopus,, Current Controlled Trials and WHO International Clinical Trials Registry Platform to identify randomized controlled trials comparing single versus sequential media for blastocyst culture and ongoing pregnancy rate...
October 2016: Journal of Assisted Reproduction and Genetics
Antonio Capalbo, Valeria Romanelli, Danilo Cimadomo, Laura Girardi, Marta Stoppa, Lisa Dovere, Domenico Dell'Edera, Filippo Maria Ubaldi, Laura Rienzi
For an IVF clinic that wishes to implement preimplantation genetic diagnosis for monogenic diseases (PGD) and for aneuploidy testing (PGD-A), a global improvement is required through all the steps of an IVF treatment and patient care. At present, CCS (Comprehensive Chromosome Screening)-based trophectoderm (TE) biopsy has been demonstrated as a safe, accurate and reproducible approach to conduct PGD-A and possibly also PGD from the same biopsy. Key challenges in PGD/PGD-A implementation cover genetic and reproductive counselling, selection of the most efficient approach for blastocyst biopsy as well as of the best performing molecular technique to conduct CCS and monogenic disease analysis...
July 16, 2016: Journal of Assisted Reproduction and Genetics
Demián Glujovsky, Cynthia Farquhar
ET is a critical step in an assisted reproduction cycle. Over the past decade there has been an increasing trend to extending culture from cleavage-stage to blastocyst transfer. There has also been a trend to single ET and reporting the success of an assisted reproductive cycle as a cumulative live-birth rate after using both fresh and frozen embryos. There is low evidence that fresh blastocyst transfer is associated with improved live-birth rates compared with fresh cleavage-stage embryos. However, in the few studies that report cumulative pregnancy rates after fresh and frozen transfers, no significant difference was found...
August 2016: Fertility and Sterility
N Gatimel, M Z Lacroix, S Chanthavisouk, N Picard-Hagen, V Gayrard, J Parinaud, R D Léandri
STUDY QUESTION: Do the embryo culture media and plastic materials used during assisted reproductive technology (ART) laboratory procedures expose embryos to bisphenol A (BPA)? SUMMARY ANSWER: BPA was not detected in embryo culture media or protein supplements at concentrations above those encountered in normal patient serum and follicular fluids. WHAT IS KNOWN ALREADY: BPA is strongly suspected of altering the epigenome during mammalian development...
July 2016: Human Reproduction
Yohan Kim, Karla C Williams, Carson T Gavin, Emily Jardine, Ann F Chambers, Hon S Leong
Cancer cell 'invasiveness' is one of the main driving forces in cancer metastasis, and assays that quantify this key attribute of cancer cells are crucial in cancer metastasis research. The research goal of many laboratories is to elucidate the signaling pathways and effectors that are responsible for cancer cell invasion, but many of these experiments rely on in vitro methods that do not specifically simulate individual steps of the metastatic cascade. Cancer cell extravasation is arguably the most important example of invasion in the metastatic cascade, whereby a single cancer cell undergoes transendothelial migration, forming invasive processes known as invadopodia to mediate translocation of the tumor cell from the vessel lumen into tissue in vivo...
May 2016: Nature Protocols
Céline Bouillon, Roger Léandri, Laurent Desch, Alexandra Ernst, Céline Bruno, Charline Cerf, Alexandra Chiron, Céline Souchay, Antoine Burguet, Clément Jimenez, Paul Sagot, Patricia Fauque
In animal studies, extensive data revealed the influence of culture medium on embryonic development, foetal growth and the behaviour of offspring. However, this impact has never been investigated in humans. For the first time, we investigated in depth the effects of embryo culture media on health, growth and development of infants conceived by In Vitro Fertilization until the age of 5 years old. This single-centre cohort study was based on an earlier randomized study. During six months, in vitro fertilization attempts (No...
2016: PloS One
M Durand, N Sermondade, C Herbemont, J Benard, H Gronier, J Boujenah, I Cédrin-Durnerin, C Poncelet, M Grynberg, C Sifer
OBJECTIVE: The aim of this study was to compare embryo development cultured in two single-step media commercially available: Fert/Sage One Step® (Origio) and Continuous Single Culture® (CSC) (Irvine Scientific). METHODS: A prospective auto-controlled study of sibling oocytes from women undergoing conventional in vitro fertilization (IVF) or intracytoplasmic sperm injection (ICSI) was performed in our center from February to June 2015. After fertilization, for every patient, half of oocytes were cultured in the single-step Fert/Sage One Step® (serie SAGE) and the other half in the single-step CSC®(serie CSC)...
March 2016: Gynécologie, Obstétrique & Fertilité
Nuno Costa-Borges, Marta Bellés, Marcos Meseguer, Daniela Galliano, Agustin Ballesteros, Gloria Calderón
OBJECTIVE: To evaluate the efficiency of using a continuous (one-step) protocol with a single medium for the culture of human embryos in a time-lapse incubator (TLI). DESIGN: Prospective cohort study on sibling donor oocytes. SETTING: University-affiliated in vitro fertilization (IVF) center. PATIENT(S): Embryos from 59 patients. INTERVENTION(S): Culture in a TLI in a single medium with or without renewal of the medium on day-3...
March 2016: Fertility and Sterility
Tai-lang Yin, Yi Zhang, Sai-jiao Li, Meng Zhao, Jin-li Ding, Wang-ming Xu, Jing Yang
Whether the type of culture media utilized in assisted reproductive technology has impacts on laboratory outcomes and birth weight of newborns in in-vitro fertilization (IVF)/intracytoplasmic sperm injection (ICSI) was investigated. A total of 673 patients undergoing IVF/ICSI and giving birth to live singletons after fresh embryo transfer on day 3 from Jan. 1, 2010 to Dec. 31, 2012 were included. Three types of culture media were used during this period: Quinn's Advantage (QA), Single Step Medium (SSM), and Continuous Single Culture medium (CSC)...
December 2015: Journal of Huazhong University of Science and Technology. Medical Sciences
Thorir Hardarson, Mona Bungum, Joe Conaghan, Marius Meintjes, Samuel J Chantilis, Laszlo Molnar, Kristina Gunnarsson, Matts Wikland
OBJECTIVE: To study whether a culture medium that allows undisturbed culture supports human embryo development to the blastocyst stage equivalently to a well-established sequential media. DESIGN: Randomized, double-blinded sibling trial. SETTING: Independent in vitro fertilization (IVF) clinics. PATIENT(S): One hundred twenty-eight patients, with 1,356 zygotes randomized into two study arms. INTERVENTION(S): Embryos randomly allocated into two study arms to compare embryo development on a time-lapse system using a single-step medium or sequential media...
December 2015: Fertility and Sterility
Jingchen Chen, Andrea Streit
BACKGROUND: Enhancers are key elements to control gene expression in time and space and thus orchestrate gene function during development, homeostasis, and disease. Whole genome approaches and bioinformatic predictions have generated a tremendous pool of potential enhancers, however their spatiotemporal activity often remains to be validated in vivo. Despite recent progress in developing high throughput strategies for enhancer evaluation, these remain mainly restricted to invertebrates and in vitro cell culture...
October 2015: Developmental Dynamics: An Official Publication of the American Association of Anatomists
Ljiljana Milenkovic, Lucien E Weiss, Joshua Yoon, Theodore L Roth, YouRong S Su, Steffen J Sahl, Matthew P Scott, W E Moerner
Accumulation of the signaling protein Smoothened (Smo) in the membrane of primary cilia is an essential step in Hedgehog (Hh) signal transduction, yet the molecular mechanisms of Smo movement and localization are poorly understood. Using ultrasensitive single-molecule tracking with high spatial/temporal precision (30 nm/10 ms), we discovered that binding events disrupt the primarily diffusive movement of Smo in cilia at an array of sites near the base. The affinity of Smo for these binding sites was modulated by the Hh pathway activation state...
July 7, 2015: Proceedings of the National Academy of Sciences of the United States of America
Fatemeh Salabi, Mahmood Nazari, Wen G Cao
BACKGROUND: This study was performed to describe the basic methods to isolate and culture of primary satellite cells (PSCs) obtained from 50 to 60-day-old sheep fetuses, single cell cloning of transfected PSCs and sexing of ovine PSCs based on the ZFY/ZFX, amelogenin and high-motility-group (HMG) box sequences. RESULTS: Three-step enzymatic digestion method increased PSCs isolation from tissue and reduced the damage of cells during long time incubation with enzymes...
December 2014: Journal of Biological Research
Hong Zhang, Qinglin Han, Xianqiang Ping, Li Li, Chong Chang, Ze Chen, Yuelong Shu, Ke Xu, Bing Sun
Vaccination is the most effective way to prevent and control infection by influenza viruses, and a cell-culture-based vaccine production system is preferred as the future choice for the large-scale production of influenza vaccines. As one of the WHO-recommended cell lines for producing influenza vaccines, Vero cells do not efficiently support the growth of the current influenza A virus vaccine donor strain, the A/Puerto Rico/8/1934 (PR8) virus. In this study, a single mutation of K86R in the NS2 protein can sufficiently render the high-yielding property to the PR8 virus in Vero cells...
August 2015: Virology
Jason E Swain
A large contributor to success during in vitro fertilization (IVF) lies in the processes occurring within the IVF laboratory. These processes make up the "culture system." This system entails numerous procedures and technical steps that must be optimized to produce a competent embryo. Notably, variations exist between programs that include differences in patient population, clinical stimulation, and other factors. Thus, a single "optimal" culture system to be utilized between all laboratories is likely not feasible...
March 2015: Seminars in Reproductive Medicine
Dorit C Kieslinger, Zhenxia Hao, Carlijn G Vergouw, Elisabeth H Kostelijk, Cornelis B Lambalk, Séverine Le Gac
OBJECTIVE: To compare the development of human embryos in microfluidic devices with culture in standard microdrop dishes, both under static conditions. DESIGN: Prospective randomized controlled trial. SETTING: In vitro fertilization laboratory. PATIENT(S): One hundred eighteen donated frozen-thawed human day-4 embryos. INTERVENTION(S): Random allocation of embryos that fulfilled the inclusion criteria to single-embryo culture in a microfluidics device (n = 58) or standard microdrop dish (n = 60)...
March 2015: Fertility and Sterility
Natsuki Hayakawa, Honami Ogoh, Mami Sumiyoshi, Yasuhisa Matsui, Saori Nishikawa, Kananko Miyamoto, Yuko Maede, Hiroshi Kiyonari, Mai Suzuki, Toshio Watanabe
ADP-ribosylation factor (Arf) 1 is thought to affect the morphologies of organelles, such as the Golgi apparatus, and regulate protein trafficking pathways. Mice have six Arf isoforms. In knockdown experiments with HeLa cells, no single Arf isoform among Arf1-5 is required for organelle morphologies or any membrane trafficking step. This suggests that the cooperation of two or more Arfs is a general feature. Although many cell biological and biochemical analyses have proven the importance of Arf1, the physiological roles of Arf1 in mice remain unknown...
October 31, 2014: Biochemical and Biophysical Research Communications
Vincent Brochard, Zichuan Liu
Nuclear transfer (NT) technique provides a powerful experimental tool to study the mechanisms of reprogramming processes and to derive NT-embryonic stem (ntES) cells from living or frozen animals. The Piezo-driven direct microinjection NT method has proved to be a valid technique to clone mice and other species. In addition, this method has been broadly used as a versatile tool for many fields of mouse micromanipulation. This chapter describes the "one step method" protocol of nuclear transfer in mouse, which combines injection of a donor cell nucleus and enucleation of MII metaphase in a single manipulation procedure...
2015: Methods in Molecular Biology
Pietro Laneve, Angela Giangrande
The last decades have witnessed the explosion of scientific interest around gene expression control mechanisms at the RNA level. This branch of molecular biology has been greatly fueled by the discovery of noncoding RNAs as major players in post-transcriptional regulation. Such a revolutionary perspective has been accompanied and triggered by the development of powerful technologies for profiling short RNAs expression, both at the high-throughput level (genome-wide identification) or as single-candidate analysis (steady state accumulation of specific species)...
2014: Journal of Visualized Experiments: JoVE
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