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Embryo culture

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https://www.readbyqxmd.com/read/28811525/totipotency-segregates-between-the-sister-blastomeres-of-two-cell-stage-mouse-embryos
#1
E Casser, S Israel, A Witten, K Schulte, S Schlatt, V Nordhoff, M Boiani
Following fertilization in mammals, it is generally accepted that totipotent cells are exclusive to the zygote and to each of the two blastomeres originating from the first mitotic division. This model of totipotency was inferred from a minority of cases in which blastomeres produced monozygotic twins in mice. Was this due to experimental limitation or biological constraint? Here we removed experimental obstacles and achieved reliable quantification of the prevalence of dual totipotency among mouse two-cell stage blastomeres...
August 15, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28811097/type-i-collagen-promotes-primary-cilia-growth-through-down-regulating-hdac6-mediated-autophagy-in-confluent-mouse-embryo-fibroblast-3t3-l1-cells
#2
Qian Xu, Weiwei Liu, Xiaoling Liu, Wuxiyar Otkur, Toshihiko Hayashi, Masayuki Yamato, Hitomi Fujisaki, Shunji Hattori, Shin-Ichi Tashiro, Takashi Ikejima
Primary cilia are microtubule-based organelles that extend from nearly all vertebrate cells. Abnormal ciliogenesis and cilia length are suggested to be associated with hypertension and obesity as well as diseases such as Meckel-Gruber syndrome. Extracellular matrix (ECM), comprising cellular microenvironment, influences cell shape and proliferation. However, influence of ECM on cilia biogenesis has not been well studied. In this study we examined the effects of type I collagen (col I), the major component of ECM, on primary cilia growth...
August 12, 2017: Journal of Bioscience and Bioengineering
https://www.readbyqxmd.com/read/28810862/the-cytotoxic-mechanism-of-epigallocatechin-gallate-on-proliferative-hacat-keratinocytes
#3
Yu-Wen Chu, Shu-Ting Liu, Ya-Lan Yang, Shih-Ming Huang, Wei-Ming Wang
BACKGROUND: Epigallocatechin gallate (EGCG) is the major ingredient of sinecatechins ointment, approved for the treatment of external genital and perianal warts. However, the molecular mechanism for EGCG's effect on warts resulting from the human papillomavirus (HPV) infection of keratinocytes is not well understood. HPV may survive in proliferative keratinocytes and may be involved in cell cycle regulation and progression. The objective of this study was to investigate the mechanism underlying EGCG's treatment on external genital warts of HPV infection through the cultured keratinocyte cells from the HaCaT cell line...
August 15, 2017: Journal of Biomedical Science
https://www.readbyqxmd.com/read/28810691/improvement-of-implantation-potential-in-mouse-blastocysts-derived-from-ivf-by-combined-treatment-with-prolactin-epidermal-growth-factor-and-4-hydroxyestradiol
#4
Miki Takeuchi, Misato Seki, Etsuko Furukawa, Akihito Takahashi, Kyosuke Saito, Mitsuru Kobayashi, Kenji Ezoe, Emiko Fukui, Midori Yoshizawa, Hiromichi Matsumoto
STUDY QUESTION: Can supplementation of medium with prolactin (PRL), epidermal growth factor (EGF) and 4-hydroxyestradiol (4-OH-E2) prior to embryo transfer improve implantation potential in mouse blastocysts derived from IVF? SUMMARY ANSWER: Combined treatment with PRL, EGF and 4-OH-E2 improves mouse blastocyst implantation rates, while alone, each factor is ineffective. WHAT IS KNOWN ALREADY: Blastocyst dormancy during delayed implantation caused by ovariectomy is maintained by continued progesterone treatment in mice, and estrogen injection rapidly activates blastocysts to implantation-induced status in vivo...
August 1, 2017: Molecular Human Reproduction
https://www.readbyqxmd.com/read/28809031/an-early-chick-embryo-culture-device-for-extended-continuous-observation
#5
Hans-Georg Sydow, Tobias Pieper, Christoph Viebahn, Nikoloz Tsikolia
Appropriate mechanical tension of the vitelline membrane as the culture substrate for the early chick embryo is frequently reported to be required for successful in vitro development. Here we describe a modified device, made of anodized aluminum, for in vitro culture which we used for studies of left-right symmetry breaking with emphasis on morphology and gene expression as readouts. The technique allows for easy, high-throughput tissue handling and provides a suitable tension in a stable and easily reproducible manner proven to be suitable for correct molecular left-right patterning and heart looping after long-term culture...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28809027/in-vitro-and-ex-ovo-culture-of-reptilian-and-avian-neural-progenitor-cells
#6
Wataru Yamashita, Toyo Shimizu, Tadashi Nomura
Reptiles and birds have been highlighted as excellent experimental models for the study of developmental biology; however, due to technical limitations in cellular analysis, dynamics of neural stem/progenitor cells of these animals remain unclear. In this chapter, we introduce the protocols for neurosphere culture and ex ovo embryonic culture of developing reptilian and avian embryos, which are modified from the method originally established for rodent embryos. Applications of these techniques provide powerful strategies for the study of comparative neural development of amniotes...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28809020/in-ovo-electroporation-methods-in-chick-embryos
#7
Hidekiyo Harada, Minoru Omi, Harukazu Nakamura
To elucidate a gene function, in vivo analysis is indispensable. We can carry out gain and loss of function experiment of a gene of interest by electroporation in ovo and ex ovo culture system on early-stage and advanced-stage chick embryos, respectively. In this section, we introduce in/ex ovo electroporation methods for the development of the chick central nervous system and visual system investigation.
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28809018/fluorescent-quail-a-transgenic-model-system-for-the-dynamic-study-of-avian-development
#8
David Huss, Rusty Lansford
Real-time four-dimensional (4D, xyzt) imaging of cultured avian embryos is an ideal method for investigating the complex movements of cells and tissues during early morphogenesis. While methods that transiently label cells, such as electroporation, are highly useful for dynamic imaging, they can also be limiting due to the number and type of cells that can be effectively targeted. In contrast, the heritable, stable, and long-term expression of a fluorescent protein driven by the exogenous promoter of a transgene overcomes these challenges...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28807939/prc2-mediated-transcriptomic-alterations-at-the-embryonic-stage-govern-tumorigenesis-and-clinical-outcome-in-mycn-driven-neuroblastoma
#9
Shoma Tsubota, Satoshi Kishida, Teppei Shimamura, Miki Ohira, Satoshi Yamashita, Dongliang Cao, Shinichi Kiyonari, Toshikazu Ushijima, Kenji Kadomatsu
Pediatric cancers such as neuroblastoma are thought to involve a dysregulation of embryonic development. However, it has been difficult to identify the critical events that trigger tumorigenesis and differentiate them from normal development. In this study, we report the establishment of a spheroid culture method that enriches early-stage tumor cells from TH-MYCN mice, a preclinical model of neuroblastoma. Using this method, we found that tumorigenic cells were evident as early as day E13.5 during embryo development, when the MYC and PRC2 transcriptomes were significantly altered...
August 14, 2017: Cancer Research
https://www.readbyqxmd.com/read/28805606/detection-of-sea-type-%C3%AE-thalassemia-in-embryo-biopsies-by-digital-pcr
#10
Ta-Hsien Lee, Ya-Chiung Hsu, Chia Lin Chang
OBJECTIVE: Accurate and efficient pre-implantation genetic diagnosis (PGD) based on the analysis of single or oligo-cells is needed for timely identification of embryos that are affected by deleterious genetic traits in in vitro fertilization (IVF) clinics. Polymerase chain reaction (PCR) is the backbone of modern genetic diagnoses, and a spectrum of PCR-based techniques have been used to detect various thalassemia mutations in prenatal diagnosis (PND) and PGD. Among thalassemias, SEA-type α-thalassemia is the most common variety found in Asia, and can lead to Bart's hydrops fetalis and serious maternal complications...
August 2017: Taiwanese Journal of Obstetrics & Gynecology
https://www.readbyqxmd.com/read/28802829/embryonic-zebrafish-primary-cell-culture-for-transfection-and-live-cellular-and-subcellular-imaging
#11
Wiebke A Sassen, Franziska Lehne, Giulio Russo, Sven Wargenau, Stefan Dübel, Reinhard W Köster
Although having great potential for live cell imaging to address numerous cell biological questions with high spatial and temporal resolution, primary cell cultures of zebrafish embryos are not widely used. We present an easy-to-use protocol for preparing primary cell cultures of 2 dpf zebrafish embryos allowing for live cell imaging of fully differentiated cells such as neurons and myocytes. We demonstrate that different cell types can be identified by morphology and expression of transgenic cell type-specific fluorescent reporters and that fluorescent cells can be sorted by flow cytometry to prepare an enriched culture...
August 9, 2017: Developmental Biology
https://www.readbyqxmd.com/read/28802113/l-carnitine-enhances-oocyte-maturation-and-improves-in%C3%A2-vitro-development-of-embryos-in-dromedary-camels-camelus-dromedaries
#12
Mohamed Fathi, K H El-Shahat
The aim of the present study was to investigate the effect of L-carnitine (LC) addition during either IVM or IVC on the developmental potential of camel oocytes. In Experiment 1; camel oocytes were matured in the absence (control) or presence of different concentrations of LC (0.25 mg, 0.5 mg, 0.75 mg and 1 mg/ml) for 30 h followed by in vitro fertilization and culture up to blastocyst stage. Our results demonstrated that oocytes treated with 0.5 mg/ml LC showed higher (P < 0.05) rates of maturation (74...
August 4, 2017: Theriogenology
https://www.readbyqxmd.com/read/28800556/supplementation-of-lycopene-in-maturation-media-improves-bovine-embryo-quality-in%C3%A2-vitro
#13
M M R Chowdhury, Byung-Hyun Choi, Imran Khan, Kyeong-Lim Lee, Ayman Mesalam, Seok-Hwan Song, Lianguang Xu, Myeong-Don Joo, Fahmida Afrin, Il-Keun Kong
This study sought to modulate factors that reduce embryo quality in in vitro culture (IVC) systems. Over eight replicates, 3075 oocytes were cultured in in vitro maturation media containing various concentrations of lycopene, followed by in vitro fertilization and culture. The percentages of MII-stage oocytes, the presumptive zygotes that underwent cleavage and developed into blastocysts were significantly (P < 0.05) higher, the intracellular ROS concentrations reduced significantly (P < 0.05) in oocytes/blastocysts, TUNEL assay demonstrates reduced apoptosis and increased total cell number per blastocyst (P < 0...
August 3, 2017: Theriogenology
https://www.readbyqxmd.com/read/28800499/effects-of-preimplantation-factor-on-interleukin-6-and-prostaglandin-f2%C3%AE-and-e2-in-the-bovine-endometrium
#14
Ruth E Wonfor, Manuela Natoli, Michael T Rose, Deborah M Nash
Preimplantation factor (PIF) is a pregnancy specific peptide with immune modulatory properties exerted on the human endometrium. Viable bovine embryos secrete PIF, but its effect on the bovine endometrial immune response is unknown, both in native and inflammatory stimulated endometrial tissue. An ex vivo bovine endometrial tissue culture model was used with lipopolysaccharide (LPS) as an inflammatory stimulant. The effect of synthetic PIF (sPIF) was assessed, in three separate experiments, on the secretion or mRNA expression of essential prostaglandins and cytokines...
August 2, 2017: Theriogenology
https://www.readbyqxmd.com/read/28800244/exploiting-analysis-of-heterogeneity-to-increase-the-information-content-extracted-from-fluorescence-micrographs-of-transgenic-zebrafish-embryos
#15
Tongying Shun, Albert H Gough, Subramaniam Sanker, Neil A Hukriede, Andreas Vogt
Zebrafish embryos are a near-ideal animal model for drug discovery because of their high genetic and physiological similarity to mammals, small size, high fecundity, and optical transparency. The latter properties make zebrafish at larval stages especially suited for high-content analysis and high throughput screening (HTS). However, inherent biological complexity and the inability to screen multiple specimens in a single well present a challenge for HTS because limiting replicates and high variability often prevent assays from reaching the stringent performance criteria demanded of large-scale screening assays...
August 11, 2017: Assay and Drug Development Technologies
https://www.readbyqxmd.com/read/28799257/comparison-of-costs-associated-with-substituting-culture-medium-with-saline-solution-for-follicular-flushing-during-oocyte-retrieval
#16
Dmitry Loktionov, Oladayo Oduola, Jenny Cloherty, Sharon M Warner, Nikhil Purandare
OBJECTIVES: To compare culture medium and saline solution for follicular flushing during oocyte retrieval with assisted reproductive technologies. METHODS: The present retrospective study analyzed data collected at Galway Fertility Clinic, Galway, Ireland, between January 1, 2015, and August 31, 2016; all patients attending the clinic for in vitro fertilization/intracytoplasmic sperm injection during this period were included. Data were stratified by whether follicular flushing was performed with culture medium or saline, and the oocyte yield rate, fertilization rate, embryo utilization rate, and biochemical and clinical pregnancy rates were compared between the groups...
August 11, 2017: International Journal of Gynaecology and Obstetrics
https://www.readbyqxmd.com/read/28799183/utilization-of-porcine-in%C3%A2-vitro-produced-parthenogenetic-embryos-for-co-transfer-with-vitrified-and-warmed-embryos
#17
Yoshiyuki Nakamura, Kazuhiro Kikuchi
The present study was conducted to evaluate the possibility of using in vitro-produced parthenogenetic (PA) embryos for co-transfer with morulae that had been collected in vivo and cryopreserved. The proportion of PA blastocysts (20.5%) was higher than that of their in vitro fertilization (IVF) counterparts (16.6%). Although there were no differences in morphology or diameter between the two groups, the number of cells in early PA blastocysts after in vitro culture for 6 days was lower than for IVF blastocysts (25...
August 10, 2017: Animal Science Journal, Nihon Chikusan Gakkaihō
https://www.readbyqxmd.com/read/28797153/-chorionic-villus-cell-culture-and-karyotype-analysis-in-1-983-cases-of-spontaneous-miscarriage
#18
S M Yuan, C Liao, D Z Li, J Z Huang, S Y Hu, M Ke, H Z Zhong, C X Yi
Objective: To investigate the relationship between spontaneous miscarriage and embryonic chromosome abnormalities, and to evaluate the clinical application of karyotype analysis by chorionic villus cell culture. Methods: The chorionic villus karyotype of 1 983 cases of miscarriage from January 2010 to July 2016 in Guangzhou Women and Children's Mecical Center were analyzed retrospectively. The miscarried chorionic villi were obtained by curettage under sterilized condition. The chromosome specimens were prepared after chorionic villus cell culture...
July 25, 2017: Zhonghua Fu Chan Ke za Zhi
https://www.readbyqxmd.com/read/28797040/genome-wide-analysis-of-transcription-factors-during-somatic-embryogenesis-in-banana-musa-spp-cv-grand-naine
#19
Shivani, Praveen Awasthi, Vikrant Sharma, Navjot Kaur, Navneet Kaur, Pankaj Pandey, Siddharth Tiwari
Transcription factors BABY BOOM (BBM), WUSCHEL (WUS), BSD, LEAFY COTYLEDON (LEC), LEAFY COTYLEDON LIKE (LIL), VIVIPAROUS1 (VP1), CUP SHAPED COTYLEDONS (CUC), BOLITA (BOL), and AGAMOUS LIKE (AGL) play a crucial role in somatic embryogenesis. In this study, we identified eighteen genes of these nine transcription factors families from the banana genome database. All genes were analyzed for their structural features, subcellular, and chromosomal localization. Protein sequence analysis indicated the presence of characteristic conserved domains in these transcription factors...
2017: PloS One
https://www.readbyqxmd.com/read/28795845/analysis-of-embryo-intactness-and-developmental-potential-following-slow-freezing-and-vitrification
#20
Lan Yu, Chanwei Jia, Yonglian Lan, Rui Song, Liying Zhou, Ying Li, Yu Liang, Shuyu Wang
The aim of this study was to identify the parameters that are related to intactness and developmental potential of a day 3 embryo after warming to improve the selection criteria used to cryopreserve and transfer embryos. We also sought to compare slow freezing and vitrification methods of cryopreservation and to evaluate the viability of non-intact embryos. Embryos warmed following slow freezing (n=220) or vitrification (n=522) were divided into 3 groups according to the proportion of surviving blastomeres (I<50%; II=50-99%; and III=100%)...
August 10, 2017: Systems Biology in Reproductive Medicine
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