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embryo in vitro production

Bruno Fernandes, Teresa Matamá, Diana Guimarães, Andreia Gomes, Artur Cavaco-Paulo
Melanin quantification is reportedly performed by absorption spectroscopy, commonly at 405 nm. Here, we propose the implementation of fluorescence spectroscopy for melanin assessment. In a typical in vitro assay to assess melanin production in response to an external stimulus, absorption spectroscopy clearly overvalues melanin content. This method is also incapable of distinguishing non-melanotic/amelanotic control cells from those that are actually capable of performing melanogenesis. Therefore, fluorescence spectroscopy is the best method for melanin quantification as it proved to be highly specific and accurate, detecting even small variations in the synthesis of melanin...
October 22, 2016: Pigment Cell & Melanoma Research
Denise Laskowski, Ylva Sjunnesson, Hans Gustafsson, Patrice Humblot, Göran Andersson, Renée Båge
BACKGROUND: Insulin has been used as a stimulatory factor for in vitro cell culture since many years. Even for routine in vitro embryo production (IVP), insulin is added to the media during different steps. There is a strong difference in concentrations used in vitro compared to what is measured in vivo in follicular fluid or serum. We performed a pilot study on insulin stability to explain possible reasons for that variation. RESULTS: We measured insulin concentrations before and after bovine oocyte maturation in an experiment by using a quantitative ELISA (Mercodia bovine insulin ELISA immunoassay) and found that concentrations were stable up to 22 h of incubation...
October 20, 2016: Acta Veterinaria Scandinavica
Mario A Cepeda, Jacob J H Pelling, Caitlin L Evered, Karla C Williams, Zoey Freedman, Ioana Stan, Jessica A Willson, Hon S Leong, Sashko Damjanovski
BACKGROUND: Membrane Type-1 Matrix Metalloproteinase (MT1-MMP) is a multifunctional protease implicated in metastatic progression ostensibly due to its ability to degrade extracellular matrix (ECM) components and allow migration of cells through the basement membrane. Despite in vitro studies demonstrating this principle, this knowledge has not translated into the use of MMP inhibitors (MMPi) as effective cancer therapeutics, or been corroborated by evidence of in vivo ECM degradation mediated by MT1-MMP, suggesting that our understanding of the role of MT1-MMP in cancer progression is incomplete...
October 18, 2016: Molecular Cancer
X L Ren, P Liu, Y Lian, J Huang, X Y Zheng, Y J Zhu, J Qiao
OBJECTIVE: To compare two flexible embryo catheters and determine whether clinical outcome differs in the in vitro fertilization-embryo transfer (IVF-ET) cycles. METHODS: This prospective control study was conducted by one doctor between July 2012 and November 2013. In the study, 2 064 patients undergoing fresh embryo transfer by using IVF-ET/intracytoplasmic sperm injection (ICSI)-ET in Reproductive Medical Center of Peking University Third Hospital were recruited...
October 18, 2016: Beijing da Xue Xue Bao. Yi Xue Ban, Journal of Peking University. Health Sciences
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
Ramiro Olivera, Lucia Natalia Moro, Roberto Jordan, Carlos Luzzani, Santiago Miriuka, Martin Radrizzani, F Xavier Donadeu, Gabriel Vichera
The demand for equine cloning as a tool to preserve high genetic value is growing worldwide; however, nuclear transfer efficiency is still very low. To address this issue, we first evaluated the effects of time from cell fusion to activation (<1h, n = 1261; 1-2h, n = 1773; 2-3h, n = 1647) on in vitro and in vivo development of equine embryos generated by cloning. Then, we evaluated the effects of using different nuclear donor cell types in two successive experiments: I) induced pluripotent stem cells (iPSCs) vs...
2016: PloS One
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
Michiko Nakai, Junya Ito, Shun-Ichi Suzuki, Dai-Ichiro Fuchimoto, Shoichiro Senbon, Misae Suzuki, Junko Noguchi, Hiroyuki Kaneko, Akira Onishi, Naomi Kashiwazaki, Kazuhiro Kikuchi
In pigs, the efficiency of embryo production after intracytoplasmic sperm injection (ICSI) is still low because of frequent failure of normal fertilization, which involves formation of two polar bodies and two pronuclei. To clarify the reasons for this, we hypothesized that ICSI does not properly trigger sperm-induced fertilization events, especially intracellular Ca(2+) signaling, also known as Ca(2+) oscillation. We also suspected that the use of in vitro-matured oocytes might negatively affect fertilization events and embryonic development of sperm-injected oocytes...
September 30, 2016: Journal of Reproduction and Development
B H Kipper, J T Trevizan, J T Carreira, I R Carvalho, G Z Mingoti, M E Beletti, S H V Perri, D A Franciscato, J C Pierucci, M B Koivisto
The aim of this study was to evaluate the chromatin packing and sperm head morphometry of cryopreserved semen of Nelore bulls (Bos taurus indicus) of different ages. Furthermore, the influence of the degree of chromatin compaction on in vitro embryo production (IVP) was investigated. Forty bulls were divided into three groups: young (1.8-2 years), adult (3.5-7 years), and senile (8-14.3 years). The ejaculates were frozen according to standards established by the Artificial Insemination Center located in the Southeast of Brazil...
August 26, 2016: Theriogenology
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
Pouria HosseinNia, Mehdi Hajian, Mojtaba Tahmoorespur, Sayyed Morteza Hosseini, Somayyeh Ostadhosseini, Mohammad Reza Nasiri, Mohammad Hossein Nasr-Esfahani
BACKGROUND: Little is understood about the regulation of gene expression during early goat embryo development. This study investigated the expression profile of 19 genes, known to be critical for early embryo development in mouse and human, at five different stages of goat in vitro embryo development (oocyte, 8-16 cell, morula, day-7 blastocyst, and day 14 blastocyst). MATERIALS AND METHODS: In this experimental study, stage-specific profiling using real time-quantitative polymerase chain reaction (RT-qPCR) revealed robust and dynamic patterns of stage-specific gene activity that fall into four major clusters depending on their respective mRNA profiles...
October 2016: International Journal of Fertility & Sterility
B Merlo, E Iacono, D Bucci, M Spinaci, G Galeati, G Mari
In vitro embryo production in the horse is still not as efficient as in other species. Oxidative stress negatively affects oocyte and embryo culture. To attenuate/minimize the oxidative stress, antioxidants such as low-molecular thiol compounds can be added to culture media. Beta-mercaptoethanol (BME) has been shown to improve maturation and embryo development in different species. The aim of this study was to investigate whether the addition to maturation medium of BME at common (0.1 mM) and high (0.7 mM) concentration could improve oocyte maturation also in the horse...
September 21, 2016: Reproduction in Domestic Animals, Zuchthygiene
P Loren, C Cheuquemán, E Sánchez, J Risopatrón, M E Arias, R Felmer, R Sánchez
Short-term exposure of gametes to different types of stress might induce stress tolerance in mammalian embryos. The aim of this study was to evaluate the effect of short-term exposure of bovine mature cumulus-oocyte complex (COC) to 3-morpholinosydnonimine (SIN-1) on subsequent in vitro embryo development, embryo quality and relative gene expression. Matured COCs were incubated with SIN-1 (0, 0.1, 1, 10 and 100 μM SIN-1) for 1 hr before in vitro fertilization and zygotes were cultured until Day 7. The cleavage rate at 72 hr did not show any differences among groups...
September 20, 2016: Reproduction in Domestic Animals, Zuchthygiene
A E Velásquez, J Manríquez, F O Castro, J F Cox, Ll Rodriguez-Alvarez
Embryo splitting has been used for the production of identical twins and to increase the pregnancy rate per available embryo. Split blastocysts can develop to term; however, little is known about the impact on gene expression of split embryos, especially at the whole transcriptome level. This work was aimed to evaluate the effect of blastocyst splitting on global gene expression profile at the elongation stage. For that, split and time-matched nonsplit (control group) bovine blastocysts were transferred to a bovine recipient and recovered at Day 17 of development...
August 22, 2016: Theriogenology
Xiao-Yu Chen, Zhi-Wei Zhu, Fu-Xian Yu, Jing Huang, Xiao-Rui Hu, Jian-Zhi Pan
Genomic integration of transgene by lentiviral vector has been proved an efficient method to produce single-transgenic animals. But it failed to create multi-gene transgenic offspring. Here, we have exploited lentivirus to generate the double-transgenic piglets through the female germline. The recombinant lentivirus containing fluorescent proteins genes (DsRed1 and Venus) were injected into the perivitelline space of 2-cell stage in vitro porcine embryos. Compared to control group, there was no significantly decreased in the proportion of blastocysts, and the two fluorescent protein genes were co-expressed in almost all the injected embryos...
June 28, 2016: Animal Reproduction Science
Elizabeth G Crichton, Clara Malo, Budhan S Pukazhenthi, Peter Nagy, Julian A Skidmore
Cholesterol (cholesterol-loaded cyclodextrins: CLC) treatment of dromedary camel sperm prior to freezing enhances cryosurvival. The present study first validated the efficacy of a heterologous zona-free goat oocyte assay (n=115 oocytes) to evaluate camel sperm function in vitro (Experiment 1: n=6 bulls), then examined the effects of CLC treatment (1.5mg/mL CLC; CLC+) versus no treatment (0 CLC) of fresh (Experiment 2: n=4 bulls) and frozen-thawed (Experiment 3: n=5 bulls) camel sperm to penetrate, de-condense and form pro-nuclei in in vitro-matured goat oocytes...
August 31, 2016: Animal Reproduction Science
Qing-Yang Li, Juan Lou, Xiao-Gan Yang, Yang-Qing Lu, Sheng-Sheng Lu, Ke-Huan Lu
Improving the quality of in vitro maturated buffalo oocytes is essential for embryo production. We report here the effects on microtubules and microfilaments in oocytes and embryo development that result from treating buffalo oocytes with the phosphodiesterase 3 (PDE3) inhibitor cilostamide. Addition of 20μM or 50μM cilostamide for 24h during in vitro maturation showed no differences in the percentage of oocytes arrested at the germinal vesicle (GV) stage. When 20μM cilostamide was added to the pre-maturation culture for 6h, 12h or 24h and continued for another 24h without cilostamide, oocytes resumed meiosis, but with significantly lower (P<0...
September 6, 2016: Animal Reproduction Science
Daniel Veraguas, Paula F Gallegos, Alejandra E Velasquez, Fidel O Castro, Lleretny Rodriguez-Alvarez
In the domestic cat, the efficiency of in vitro embryo production systems is negatively affected during the nonbreeding season. The objective of this research was to evaluate the effect of FSH stimulation in anestrous cats, on quality of cumulus-oocyte complexes (COCs) and in vitro developmental competence after parthenogenetic activation. To accomplish this purpose, anestrous cats were grouped into: (1) FSH treated (serial doses of 5 mg of porcine FSH each, every 24 hours, for 4 days) and (2) untreated control...
August 13, 2016: Theriogenology
Laila Noli, Antonio Capalbo, Yaser Dajani, Danilo Cimadomo, Jean Bvumbe, Laura Rienzi, Filippo Maria Ubaldi, Caroline Ogilvie, Yacoub Khalaf, Dusko Ilic
Studies reporting term pregnancy and the production of genetically identical offspring from isolated blastomeres of early stage embryos have been carried out in small and large animals. However, very little is known about the effects of embryo splitting on the development and reproductive competency of human embryos. In this study, we investigated the effects of embryo splitting on profile of microRNAs (miRNAs) detected in their spent blastocyst medium (SBM) by comparative analysis of miRNA profiles in SBM of human twin embryos created by blastomere biopsy and SBM of blastocysts that resulted in a healthy pregnancy and live birth following embryo transfer...
October 17, 2016: Stem Cells and Development
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