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Embryo Culture (embryo + culture)
Selected AbstractsIn Vitro Production of Equine Embryos: State of the ArtREPRODUCTION IN DOMESTIC ANIMALS, Issue 2010K Hinrichs Contents In vitro embryo production is possible in the horse both clinically and for research applications. Oocytes may be collected from excised ovaries post-mortem, or from either immature follicles or stimulated pre-ovulatory follicles in the live mare. In vitro maturation of immature oocytes typically yields approximately 60% mature oocytes. As standard in vitro fertilization is not yet repeatable in the horse, fertilization is performed by intracytoplasmic sperm injection. Embryo culture requires medium with high glucose, at least during blastocyst development, and rates of blastocyst development similar to those for cattle (25% to 35%) may be obtained. Pregnancy rates after transfer of in vitro -produced blastocysts are similar to those for embryos recovered ex vivo. [source] Lack of teratogenicity of microcystin-LR in the mouse and toadJOURNAL OF APPLIED TOXICOLOGY, Issue 1 2002N. Chernoff Abstract Microcystin-LR (MC-LR) is a cyanobacterial toxin generated by the organism Microcystis aeruginosa. Although the hepatotoxicity of this chemical has been characterized, the potential developmental toxicity in vertebrates has not been well studied. The purpose of this study was to elucidate the effects of this toxin on the in vivo and in vitro development of mammals and the development of an Anuran (toad). Initial acute toxicity experiments with female CD-1 mice were accomplished with MC-LR administered i.p. in saline. Lethality occurred at 128 and 160 µg kg ,1 and histopathology revealed massive hepatic necrosis with diffuse hemorrhage. Developmental toxicity studies were done with MC-LR administered i.p. for 2-day periods: gestation days 7,8, 9,10 or 11,12. Doses used ranged from 2 to 128 µg kg,1. On gestation day 17, fetuses were weighed and analyzed for gross morphological and skeletal defects. No treatment-related differences were seen in litter size, viability, weight or the incidence of anomalies. Groups of dams dosed with 32,128 µg kg,1 on gestation days 7,8, 9,10 or 11,12 were allowed to give birth and the growth and development of their pups were followed postnatally. There were no significant effects noted in the offspring of the treated dams. Neurulation-staged CD-1 mouse conceptuses were exposed to 50,1000 nM MC-LR in whole embryo culture for 24 h. No significant increase in abnormalities or developmental delays was observed. Finally, exposure of the developing toad. Bufo arenarum was done from stage 17 (tail bud) for 10 days at concentrations of 1,20 mg l,1. No effect on morphological development or survival was noted in any exposed groups. These data indicate that microcystin does not appear to affect development adversely in the mouse (in vivo or in vitro) or the toad at the doses and exposure parameters used. Copyright © 2002 John Wiley & Sons, Ltd. [source] Characterization of an immortalized oviduct cell line from the cynomolgus monkey (Macaca fascicularis)JOURNAL OF MEDICAL PRIMATOLOGY, Issue 2 2005H. Okada Abstract:, To establish reproductive biological techniques in mammals, it is important to understand the growth environment of the embryo. Oviduct epithelial cells are in close proximity to the embryo during pre-implantation development. We, therefore, established an immortalized oviduct epithelial cell line from the cynomolgus monkey, evaluated the usefulness of these cells as feeder cells for embryo culture, and investigated the gene expression of several growth factors and cytokines in the cells. The immortalized cells were positive for the anti-cytokeratin antibody, as determined by immunocytochemistry, indicating that they are epithelial. They also expressed oviductin, which is specific to oviduct epithelial cells, glyceraldehyde-3-phosphate dehydrogenase (control), leukemia inhibitory factor, vascular endothelial growth factor, epidermal growth factor, insulin-like growth factor 1, transforming growth factor beta-2, and interleukin 4. Mouse embryo development was improved when the immortalized cells were used as feeder cells. This cell line is also useful for studying the factors secreted by oviduct epithelial cells. [source] Quantifying oxygen diffusion in paraffin oil used in oocyte and embryo cultureMOLECULAR REPRODUCTION & DEVELOPMENT, Issue 12 2009Yvonne M. StokesArticle first published online: 11 AUG 200 Oxygen diffusion through oil is important in the culture of oocytes and embryos. A diffusion coefficient two orders of magnitude smaller than that of oxygen in water has been thought possible, and this has led to concerns of anoxia in cultures. Using an assay for determining the oxygen consumption rate of embryos and oocytes, along with a mathematical model, it is here shown that the oxygen diffusion rate in paraffin oil at 37°C is about two-thirds of that in water at the same temperature. Although not previously recognised for the assay in question, the geometry is such that anoxia does occur for a period of time in excess of 1,hr and, by the completion of the assay, 30,40% of the medium is anoxic. Hence the quantity of oxygen consumed is less than would be consumed in conditions of plentiful oxygen supply. Nevertheless, using a model with a concentration dependent oxygen consumption rate, the oxygen consumption rate can be estimated. Mol. Reprod. Dev. 76: 1178,1187, 2009. © 2009 Wiley-Liss, Inc. [source] Actions of Tumor Necrosis Factor-, on Oocyte Maturation and Embryonic Development in Cattle,AMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY, Issue 5 2003P. Soto Problem:, Infertility can accompany mastitis in cattle. Involvement of tumor necrosis factor- , (TNF- ,) in this phenomenon is suggested by observations that circulating concentrations of TNF- , are elevated after intramammary infection or infusion of endotoxin. It was hypothesized that (1) TNF- , acts on the oocyte during maturation to decrease the percent of oocytes that cleave and develop following fertilization; (2) exposure of embryos to TNF- , after fertilization reduces development to the blastocyst stage; and (3) TNF- , increases the proportion of blastomeres that undergo apoptosis in a stage-of-development dependent manner. Method of study:, In one experiment, oocytes were matured with various concentrations of TNF- , and then fertilized and cultured without TNF- ,. In another study, embryos were cultured with TNF- , for 8 days beginning after fertilization. Finally, embryos were collected at the two or four-cell stage (at 28,30 hr after insemination) or when ,9-cells (at day 4 after insemination) and cultured ± TNF- , for 24 hr. The proportion of blastomeres undergoing apoptosis was then determined by the TUNEL procedure. Results:, Addition of TNF- , to maturation medium did not affect the proportion of oocytes that cleaved. However, the percent of oocytes that developed to the blastocyst stage at day 8 after insemination was reduced (P = 0.05) at all TNF- , concentrations tested (0.1,100 ng/mL). When added during embryo culture, there was no significant effect of TNF- , on the proportion of oocytes that became blastocysts. In addition, TNF- , did not induce apoptosis in two and four-cell embryos. For embryos ,9-cells, however, 10 and 100 ng/mL TNF- , increased (P < 0.05) the percent of blastomeres labeling as TUNEL-positive. Conclusion:, TNF- , can have deleterious actions on oocyte maturation that compromise development of the resultant embryo. While exposure of fertilized embryos to TNF- , did not inhibit development to the blastocyst stage, TNF- , increased the percentage of blastomeres undergoing apoptosis when exposure occurred for embryos ,9-cells. Increased blastomere apoptosis could conceivably compromise subsequent embryo survival. [source] Effect of Anti-Basic Fibroblast Growth Factor (Anti-bFGF) on In Vitro Embryonic Development in RatANATOMIA, HISTOLOGIA, EMBRYOLOGIA, Issue 4 2009E. Unur Summary In this study, we aimed at the in vitro effects of anti-fibroblast growth factor-2 (anti-FGF-2 or anti-bFGF) on embryo culture in rats. In vitro effects of anti-bFGF on total embryonic development were investigated in 40 rat embryos (which were divided into four groups) (obtained from five pregnant females) at 9.5 days of gestation that were cultured in whole rat serum (WRS), and in WRS+ 2.5, 5, and 10 ,g/ml anti-bFGF. After 48 h of culturing, the embryos from each group were harvested to be analysed morphologically according to a morphological scoring system and biochemically to obtain the embryo protein content. The morphological score, embryo protein content, somite number and crown-rump length of embryos indicated that embryos cultured in WRS+ anti-bFGF had significant embryonic retardation. Mean morphological scores for the embryos grown in WRS, in the presence of 2.5, 5 and 10 ,g anti-FGF-2 were 61.4 ± 1.64, 46.3 ± 8.42, 27 ± 2.58 and13.6 ± 0.96 respectively. These results suggest that bFGF is very important for normal embryonic development and rat anti-bFGF neutralizes bFGF effect. [source] Investigation of Direct Toxic and Teratogenic Effects of Anticoagulants on Rat Embryonic Development Using In Vitro Culture Method and Genotoxicity AssayANATOMIA, HISTOLOGIA, EMBRYOLOGIA, Issue 2 2006I. I. Uysal Summary Heparin and low molecular weight heparins (LMWHs) are used to reduce the incidence of venous thromboembolism in pregnancy. Although, these agents have been shown to be safe when used during pregnancy, the studies about direct toxic and teratogenic effects of these drugs on embryonic development are limited. In this study, the effects of heparin and LMWHs on rat embryonic development were investigated by using in vitro embryo culture and micronucleus (MN) assay methods. Rat embryos were cultured in vitro in the presence of different concentrations of heparin (5,40 IU/ml), dalteparin (2.5,20 IU/ml), enoxaparin (25,100 ,g/ml) and nadroparin (1,4 IU/ml). Effects of anticoagulants on embryonic developmental parameters were compared and embryos were evaluated for the presence of any malformations. After culturing the embryos, classic MN assay was performed. Anticoagulants significantly decreased all growth and developmental parameters dose-dependently. Dalteparin and enoxaparin were found to cause more developmental toxicity than heparin and nadroparin. Along with haematoma in general, heparin and nadroparin caused maxillary deformity, situs inversus and oedema most frequently, while neural tube defects were observed with dalteparin and enoxaparin. All agents also significantly induced MN formation in rat embryonic blood cells. These results indicate the possible genotoxic effects of anticoagulant agents on the developing rat embryo when applied directly. [source] Developmental toxicity of methanol: Pathogenesis in CD-1 and C57BL/6J mice exposed in whole embryo culture,,BIRTH DEFECTS RESEARCH, Issue 4 2004Sigmund J. Degitz BACKGROUND Methanol causes axial skeleton and craniofacial defects in both CD-1 and C57BL/6J mice during gastrulation, but C57BL/6J embryos are more severely affected. We evaluated methanol-induced pathogenesis in CD-1 and C57BL/6J embryos exposed during gastrulation in whole embryo culture. METHODS Conceptuses with five to seven somites were exposed to 0, 1, 2, 3, 4, or 6 mg methanol/ml culture medium for 24 hr and embryonic morphology was assessed. Cell death was evaluated by histology and LysoTracker red staining, and cell-cycle distribution was evaluated by flow cytometry. RESULTS In C57BL/6J embryos, craniofacial defects were observed at 3 mg methanol/ml and greater. The response for CD-1 embryos was different, with increased dysmorphology only at 6 mg/ml. However, protein content in CD-1 embryos was reduced at 3 mg methanol/ml and above, indicating growth retardation. Yolk sac toxicity occurred only at 6 mg methanol/ml in both strains. Methanol caused only small changes in cell-cycle distribution, while cell death was induced at 4 and 6 mg methanol/ml in both strains after 8 hr. The extent of cell death after 8 hr was greater in C57BL/6J embryos, and increased over time through 18 hr; in contrast, CD-1 embryos showed less cell death at 18 than at 8 hr, suggesting recovery. CONCLUSIONS Cell death plays a prominent role in methanol-induced dysmorphogenesis, while cell-cycle perturbation may not. Differences in the extent of cell death between CD-1 and C57BL/6J embryos correlated with differences in the severity of dysmorphogenesis. Birth Defects Research (Part A), 2004. Published 2004 Wiley-Liss, Inc. [source] | |||||||||||||