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Embryo culture

Cassiano R A F Diniz, Plinio C Casarotto, Senem M Fred, Caroline Biojone, Eero Castrén, Sâmia R L Joca
The renin-angiotensin system (RAS) is associated with peripheral fluid homeostasis and cardiovascular function, but recent evidence also suggests a functional role in the brain. RAS regulates physiological and behavioral parameters related to the stress response, including depressive symptoms. Apparently, RAS can modulate levels of brain-derived neurotrophic factor (BDNF) and TRKB, which are important in the neurobiology of depression and antidepressant action. However, the interaction between the BDNF/TRKB system and RAS in depression has not been investigated before...
March 14, 2018: Neuropharmacology
Eva Pallinger, Zoltan Bognar, Agnes Bogdan, Timea Csabai, Hajnalka Abraham, Julia Szekeres-Bartho
Earlier evidence suggests, that the embryo signals to the maternal immune system. Extracellular vesicles (EVs) are produced by all types of cells, and because they transport different kinds of molecules from one cell to the other, they can be considered as means of intercellular communication. The aim of this work was to test, whether the embryo is able to produce sufficient amounts of EVs to alter the function of peripheral lymphocytes. Embryo-derived EVs were identified by their Annexin V biding capacity, and sensitivity to Triton X dependent lysis, using flow cytometry...
March 16, 2018: Scientific Reports
T Arthur Chang, Gennadiy I Bondarenko, Behzad Gerami-Naini, Jessica G Drenzek, Maureen Durning, Mark A Garthwaite, Jenna Kropp Schmidt, Thaddeus G Golos
BACKGROUND: The initiation of primate embryo invasion into the endometrium and the formation of the placenta from trophoblasts, fetal mesenchyme, and vascular components are essential for the establishment of a successful pregnancy. The mechanisms which direct morphogenesis of the chorionic villi, and the interactions between trophectoderm-derived trophoblasts and the fetal mesenchyme to direct these processes during placentation are not well understood due to a dearth of systems to examine and manipulate real-time primate implantation...
March 16, 2018: Reproductive Biology and Endocrinology: RB&E
Jihyun Kim, Haesun A Kim
The most widely used method (Brockes' method) for preparing primary Schwann cell culture uses neonatal rat sciatic nerves as the primary source of Schwann cells. The procedure is relatively simple and yields a highly purified population of Schwann cells in a short period of time. The method has also been used to prepare Schwann cells from mice, however, with limitation. For example, Brockes' method is not applicable when the genotypes of mouse neonates are unknown or if the mouse mutants do not develop to term...
2018: Methods in Molecular Biology
Rhona Mirsky, Kristjan R Jessen
Schwann cell precursors are the first defined stage in the generation of Schwann cells from the neural crest and represent the glial cell of embryonic nerves. Highly pure cultures of these cells can be obtained by enzymatic dissociation of nerves dissected from the limbs of 14- or 12-day-old rat and mouse embryos, respectively. Since Schwann cell precursors, unlike Schwann cells, are acutely dependent on axonal signals for survival, they require addition of trophic factors, typically β-neuregulin-1, for maintenance in cell culture...
2018: Methods in Molecular Biology
Masami Hayashi, Kayo Yoshida, Kohei Kitada, Akane Kizu, Daisuke Tachibana, Mitsuru Fukui, Takashi Morita, Masayasu Koyama
PURPOSE: To study the outcomes of mouse preimplantation embryos irradiated with low doses of X-rays (≤ 1 Gy) and investigate apoptosis and pluripotency of the irradiated embryos. METHODS: Mouse embryos at the 2-cell stage were collected for in vitro culture. After reaching the 8-cell stage, embryos were irradiated with various low doses of X-rays (0-1 Gy). Blastocysts with a normal appearance were transferred into a pseudopregnant uterus. The developmental rate to blastocysts and the survival rate following embryo transfer were examined...
March 16, 2018: Journal of Assisted Reproduction and Genetics
Lanlin Yang, Sufen Cai, Shuoping Zhang, Xiangyi Kong, Yifan Gu, Changfu Lu, Jing Dai, Fei Gong, Guangxiu Lu, Ge Lin
STUDY QUESTION: Does single cleavage-stage (Day 3) embryo transfer using a time-lapse (TL) hierarchical classification model achieve comparable ongoing pregnancy rates (OPR) to single blastocyst (Day 5) transfer by conventional morphological (CM) selection? SUMMARY ANSWER: Day 3 single embryo transfer (SET) with a hierarchical classification model had a significantly lower OPR compared with Day 5 SET with CM selection. WHAT IS KNOWN ALREADY: Cleavage-stage SET is an alternative to blastocyst SET...
March 13, 2018: Human Reproduction
Panyu Chen, Tingting Li, Lei Jia, Cong Fang, Xiaoyan Liang
The purpose of this study is to determine whether all-blastocyst-culture can benefit advanced maternal age (AMA) patients with low ovarian reserve. We retrospectively analyze AMA patients with low ovarian reserve undergoing IVF/ICSI in Reproductive Medicine Research Center of the Sixth Affiliated Hospital of Sun Yat-sen University from May 2016 to August 2017. We divide them into group A (all-blastocyst-culture) and group B (none-blastocystculture) based on different culture approach of D3 transferrable embryo...
March 16, 2018: Gynecological Endocrinology
Alexandra J Harvey, Carmel O'Brien, Jack Lambshead, John R Sheedy, Joy Rathjen, Andrew L Laslett, David K Gardner
Reprogramming somatic cells to a pluripotent cell state (induced Pluripotent Stem (iPS) cells) requires reprogramming of metabolism to support cell proliferation and pluripotency, most notably changes in carbohydrate turnover that reflect a shift from oxidative to glycolytic metabolism. Some aspects of iPS cell metabolism differ from embryonic stem (ES) cells, which may reflect a parental cell memory, or be a consequence of the reprogramming process. In this study, we compared the metabolism of 3 human iPS cell lines to assess the fidelity of metabolic reprogramming...
2018: PloS One
Kathrine Vauvert R Hviid, Sara Sofia Malchau, Anja Pinborg, Henriette Svarre Nielsen
BACKGROUND: The incidence of monozygotic twins (MZT) after ART appears to be higher than the incidence after spontaneous conceptions contradicting the aim of ART to avoid multiple pregnancies because of the associated risks. OBJECTIVE AND RATIONALE: The aim was to study the frequency of MZT after IVF and ICSI and how it is influenced by the day of embryo transfer, maternal age, zona pellucida manipulation, controlled ovarian stimulation, stimulation protocol, culture media and embryo quality...
March 12, 2018: Human Reproduction Update
Ellen A G Chernoff, Kazuna Sato, Hai V N Salfity, Deborah A Sarria, Teri Belecky-Adams
The differentiated state of spinal cord ependymal cells in regeneration-competent amphibians varies between a constitutively active state in what is essentially a developing organism, the tadpole of the frog Xenopus laevis , and a quiescent, activatable state in a slowly growing adult salamander Ambystoma mexicanum , the Axolotl. Ependymal cells are epithelial in intact spinal cord of all vertebrates. After transection, body region ependymal epithelium in both Xenopus and the Axolotl disorganizes for regenerative outgrowth (gap replacement)...
2018: Frontiers in Cellular Neuroscience
Fan Wang, Xinghua Zhong, Haibin Wang, Aiping Song, Fadi Chen, Weimin Fang, Jiafu Jiang, Nianjun Teng
Most chrysanthemum cultivars are self-incompatible, so it is very difficult to create pure lines that are important in chrysanthemum breeding and theoretical studies. In our previous study, we obtained a self-compatible chrysanthemum cultivar and its self-pollinated seed set was 56.50%. It was interesting that the seed set of its ten progenies ranged from 0% to 37.23%. Examination of the factors causing the differences in the seed set will lead to an improved understanding of chrysanthemum self-incompatibility, and provide valuable information for creating pure lines...
March 13, 2018: International Journal of Molecular Sciences
R Poverini, R Lisi, F Lisi, V Berlinghieri, W Bielli, P Carfagna, A Costantino, D Iacomino, G Nicodemo
Granulocyte-macrophage colony-stimulation factor plays different crucial roles during embryo implantation and subsequent development. Here we aimed to evaluate the effects of embryo cell culture medium, with the inclusion of granulocyte-macrophage colony-stimulation factor (GM-CSF), on embryo development and pregnancy rate. To this end, we took advantage of our retrospective observational study to correlate the outcomes from two different culture media. We included in this study 25 unselected patient from our IVF Center that underwent heterologous IVF cycle with crypreserved oocytes...
December 2018: Cell Death Discovery
Jun Jiang, Shulin Tang, Jianhong Xia, Jikai Wen, Shuang Chen, Xiaodong Shu, Michael S Y Huen, Yiqun Deng
Wnt/β-catenin signaling activity is maintained in homeostasis by an expanding list of molecular determinants. However, the molecular components and the regulatory mechanisms involved in its fine-tuning remain to be determined. Here, we identified C9orf140, a tumor-specific protein, as a novel Axin1-interacting protein by tandem-affinity purification and mass spectrometry. We further showed that C9orf140 is a negative regulator of Wnt/β-catenin signaling in cultured cells as well as in zebrafish embryos. It functions upstream of β-catenin, outcompetes PP2A for binding to Axin1, influences the balance between phosphorylation and de-phosphorylation of β-catenin, and ultimately compromises Wnt3A-induced β-catenin accumulation...
March 13, 2018: Oncogene
Sébastien Bélanger, Suzanne Marchand, Pierre-Étienne Jacques, Blake Meyers, François Belzile
In barley, it is possible to induce embryogenesis in the haploid and uninucleate microspore to obtain a diploid plant that is perfectly homozygous. To change developmental fates in this fashion, microspores need to engage in cellular de-differentiation, interrupting the pollen formation, and restore totipotency prior to engaging in embryogenesis. In this work, we used the barley cultivar Gobernadora to characterize the transcriptome of microspores prior to (day 0) and immediately after (days 2 and 5) the application of a stress pretreatment...
March 12, 2018: G3: Genes—Genomes—Genetics
E Mangoli, M A Khalili, A R Talebi, S Ghasemi-Esmailabad, A Hosseini
The main goal was to evaluate the correlation between sperm parameters and chromatin quality with embryo kinetics via time-lapse monitoring system (TLM). A total of 40 couples involved in the ICSI program as a result of male infertility. For assessment of sperm chromatin and DNA quality, we used aniline blue, toluidine blue, chromomycin A3, acridine orange and terminal transferase-mediated deoxyuridine triphosphate biotin end labelling assays. All mature oocytes were injected, and the generated zygotes (2PNs) were cultured in TLM...
March 12, 2018: Andrologia
Kylie Beck, Jaswant Singh, Mohammad Arshud Dar, Muhammad Anzar
BACKGROUND: A suitable culture system is important for follicle growth in adult bovine ovarian tissue. This study aimed to assess the avian chorioallantoic membrane (CAM) for short-term culture of adult bovine ovarian tissues compared with a traditional in vitro culture system. METHODS: Ovarian cortical tissues (1-2 mm3 ), collected from slaughtered adult cows, were randomly assigned to control, CAM or in vitro culture groups. In the control group, ovarian tissues were fixed with paraformaldehyde without culture...
March 9, 2018: Reproductive Biology and Endocrinology: RB&E
Simon Menanteau-Ledouble, Mark L Lawrence, Mansour El-Matbouli
BACKGROUND: Like many members of the Enterobacteriaceae family, Yersinia ruckeri has the ability to invade non professional phagocytic cells. Intracellular location is advantageous for the bacterium because it shields it from the immune system and can help it cross epithelial membranes and gain entry into the host. In the present manuscript, we report on our investigation regarding the mechanisms of Y. ruckeri's invasion of host cells. RESULTS: A gentamycin assay was applied to two isolates, belonging to both the biotype 1 (ATCC 29473) and biotype 2 (A7959-11) and using several cell culture types: Atlantic Salmon Kidney, Salmon Head Kidney and, Chinook salmon embryos cells at both low and high passage numbers...
March 9, 2018: BMC Veterinary Research
Kristin M Whitworth, Raissa Cecil, Joshua A Benne, Bethany K Redel, Lee D Spate, Melissa S Samuel, Randall S Prather, Kevin D Wells
Genetically engineered pigs are often created with a targeting vector that contains a loxP flanked selectable marker like neomycin. The Cre-loxP recombinase system can be used to remove the selectable marker gene from the resulting offspring or cell line. Here is described a new method to remove a loxP flanked neomycin cassette by direct zygote injection of an mRNA encoding Cre recombinase. The optimal concentration of mRNA was determined to be 10 ng/μL when compared to 2 and 100 ng/μL (P < 0.0001)...
March 7, 2018: Transgenic Research
Ellen Moura Vale, Ricardo Souza Reis, Lucas Zanchetta Passamani, Claudete Santa-Catarina, Vanildo Silveira
Efficient protocols for somatic embryogenesis of papaya ( Carica papaya L.) have great potential for selecting elite hybrid genotypes. Addition of polyethylene glycol (PEG), a nonplasmolyzing osmotic agent, to a maturation medium increases the production of somatic embryos in C . papaya . To study the effects of PEG on somatic embryogenesis of C . papaya , we analyzed somatic embryo development and carbohydrate profile changes during maturation treatments with PEG (6%) or without PEG (control). PEG treatment (6%) increased the number of normal mature somatic embryos followed by somatic plantlet production...
March 2018: Physiology and Molecular Biology of Plants: An International Journal of Functional Plant Biology
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