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Journal ArticleDOI

Preimplantation development of in vitro-matured and in vitro-fertilized ovine zygotes: comparison between coculture on oviduct epithelial cell monolayers and culture under low oxygen atmosphere.

TL;DR: Two distinct culture environments are characterized, each capable of supporting the development of high frequencies of unselected IVMF zygotes to the blastocyst stage in vitro, and survival of the embryo under a reduced oxygen atmosphere is investigated.
Abstract: The roles of medium composition, serum source, embryo coculture, and culture under low O2 conditions on the development of in vitro-matured and in vitro-fertilized (IVMF) ovine zygotes were investigated in three separate experiments. In the first experiment, the proportion of cocultured IVMF zygotes developing to the blastocyst stage was significantly higher (38.0% vs. 3.5%; p < 0.05) than that of non-cocultured zygotes treated within three embryo culture media (TCM-199 + 10% fetal bovine serum [FBS]; bicarbonate-buffered, glucose-free synthetic oviduct fluid medium [mod-SOFM] + 10% FBS; and bicarbonate-buffered BSA-free Tyrode's salt solution [mod-TALP] + 10% FBS) under a 5% CO2 atmosphere in air. In a second experiment, a significantly higher (p < 0.05) proportion of cocultured zygotes placed in TCM-199 medium survived to the blastocyst stage (37.4% blastocysts vs. 23.4% in mod-SOFM). No significant effect of serum (FBS vs. human serum [HS]) was observed on embryonic development, but coculture was confirmed to exert a significant influence on development to the blastocyst stage. In the final experiment, survival of the embryo under a reduced oxygen (5% CO2:5% O2:90% N2) atmosphere was investigated. In contrast to results in the initial experiments, embryonic survival was significantly higher (p < 0.05) in the non-cocultured treatment groups (21.9% blastocysts vs. 0.4% for cocultured zygotes). Serum source also had a significant (p < 0.05) influence upon the development of non-cocultured zygotes: 32.3% of zygotes cultured with HS progressed to the blastocyst stage vs. 11.5% of zygotes cultured in FBS-supplemented medium. These results have characterized two distinct culture environments, each capable of supporting the development of high frequencies of unselected IVMF zygotes to the blastocyst stage in vitro.

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Citations
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Journal ArticleDOI
TL;DR: Mechanisms underlying the adaptation of pig embryos to oxygen deficiency in the maternal-conceptus microenvironment are determined and understating of cellular adaptation of developing pre-implantation porcine conceptuses to hypoxic stress is advanced.

8 citations

01 Jan 2000
TL;DR: This document summarizes current capabilities, research and operational priorities, and plans for further studies that were established at the 2015 USGS workshop on quantitative hazard assessments of earthquake-triggered landsliding and liquefaction.
Abstract: ....................................................................................................................................v

8 citations

Journal ArticleDOI
TL;DR: These are the first primate studies showing detrimental effects of reduced-oxygen culture on in vitro maturation and maturation was enhanced with complex high-glucose medium suggesting that the predominant metabolism is aerobic glycolysis.
Abstract: Purpose:The objectives of these in vitro maturation studies in primate cumulus-oocyte complexes (COCs) were to evaluate the effect of a reduced-oxygen environment and to compare medium with a high-glucose concentration to medium with pyruvate but no glucose.

8 citations


Cites background from "Preimplantation development of in v..."

  • ...Metabolism in early embryos is primarily oxidative, and they have been shown to benefit from culture in a reducedoxygen environment (21,39)....

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Dissertation
01 Jan 2012
TL;DR: For mouse study, embryonic development can be achieved from Heparin-sperm capacitation treatment, using any sperm movement for 13 to 15 hours of post-hCG suntikan oocytes, and for goat study, LOPU is good procedure to provide consistent high quality oocytes.
Abstract: Selected sperm and oocyte factors related were evaluated in ICSI experiments involving mouse and goat species in the present study. The mouse oocyte collection was conducted through oviduct retrieval, while the goat oocytes were retrieved via LOPU technique. The effectiveness of sperm capacitation chemical treatment (Heparin and Theophylline) for 1 hour and sperm movement (Rapid and Slow) were identified in both species. For oocyte factors, the effects of post-hCG injection durations (13-15 hours and 16-18 hours) in mice were studied. LOPU cycles (OR1, OR2 and OR3) and IVM durations (18-21 hours and 22-25 hours) were investigated in goats. In Experiments 1, the effects of different sperm capacitation chemicals and sperm movement of mouse sperm were studied. Mouse ICSI-embryonic development at early stages (2- and 4-cell stage) were improved by using Heparin compared to Theophylline (64.67±1.69% versus 60.50±1.21% and 50.55±2.01% versus 45.09±1.60%, respectively). Both sperm movement factors had same potential to produce mouse ICSI-derived embryos. In Experiments 2, the effect of post-hCG injection duration in mouse oocytes on embryonic development rates were studied. Two different durations were significantly different (P<0.05) on all cleavage rates (2-, 4-, 8-cell and morula stage) whereby at 13-15 hours were higher than at 16-18 hours duration (67.95±1.06% versus 57.02±1.33%, 53.51±1.13% versus 41.90±1.98%, 39.40±1.01% versus 22.21±1.62%, and 14.83±1.09% versus 9.77±1.02%, respectively). Experiment 3 evaluated the effect of two different sperm capacitation chemicals (Heparin and Theophylline) and sperm movement (Rapid and Slow) of goat sperm were studied. Both sperm capacitation chemicals gave same potential of goat ICSI-embryo iv development (P>0.05). Rapid sperm movement gave better goat ICSI-embryonic development (4-, 8- cell and morula stage), whereby the respective values were 56.62±4.69% versus 41.80±4.49%, 39.51±4.70% versus 24.24±4.05% and 9.21±2.85% versus 2.24±0.94%. Experiment 4 investigated the effect of LOPU cycles on the yield of different oocyte grades (Grades A, B, C, D and E) from 16 donors goat. Lower number of Grades D and E (31 and 19, respectively) oocytes were obtained compared to Grades A, B and C. Higher number of Grade C oocytes were obtained compared to Grades A and B from LOPU technique (153 versus 106 and 91, respectively). OR1 gave better quantity and quality oocytes, followed by OR2 and OR3 (number of oocytes per ovary were 5.47±0.67, 3.94±0.44 and 3.09±0.50, respectively). In another sub-experiment, maturation rate of goat oocytes at 18 to 21 hours was higher than 22 to 25 hours of IVM duration (97.34±7.86% versus 89.01±2.41%, respectively). However, both IVM durations had the same potential in ICSI-derived embryo development. In conclusion, for mouse study, embryonic development can be achieved from Heparin-sperm capacitation treatment, using any sperm movement for 13 to 15 hours of post-hCG suntikan oocytes. For goat study, LOPU is good procedure to provide consistent high quality oocytes (Grades A, B and C) and it usage can be repeated on the same donor up to 3 times. Rapid sperm movement is a better choice for ICSI-embryonic development. Both sperm capacitation chemical treatments have the same potential to produce ICSI-embryonic development.

7 citations


Cites background from "Preimplantation development of in v..."

  • ..., 1992), slicing the ovary (Slavik et al., 1992; Watson et al., 1994) and follicular dissection (Fukui et al....

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References
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Journal ArticleDOI
TL;DR: In this article, a general class of regression models for ordinal data is developed and discussed, which utilize the ordinal nature of the data by describing various modes of stochastic ordering and this eliminates the need for assigning scores or otherwise assuming cardinality instead of ordinality.
Abstract: SUMMARY A general class of regression models for ordinal data is developed and discussed. These models utilize the ordinal nature of the data by describing various modes of stochastic ordering and this eliminates the need for assigning scores or otherwise assuming cardinality instead of ordinality. Two models in particular, the proportional odds and the proportional hazards models are likely to be most useful in practice because of the simplicity of their interpretation. These linear models are shown to be multivariate extensions of generalized linear models. Extensions to non-linear models are discussed and it is shown that even here the method of iteratively reweighted least squares converges to the maximum likelihood estimate, a property which greatly simplifies the necessary computation. Applications are discussed with the aid of examples.

3,647 citations

Journal ArticleDOI
TL;DR: A procedure to obtain high and repeatable fertilization frequencies for bovine in vitro fertilization (IVF) with frozen-thawed sperm was developed and Heparin was the most important factor in increasing IVF frequencies.

1,165 citations

Journal ArticleDOI
TL;DR: The Two Faces of Oxygen Molecular oxygen is both benign and malign and the superoxide dismutases, by catalytically scavenging 0;, provide a defense against it and against any reactive radical species which can be derived from it.
Abstract: The Two Faces of Oxygen Molecular oxygen is both benign and malign On the one hand it provides enormous advantages and on the other it imposes a universal toxicity This toxicity is largely due to the intermediates of oxygen reduction, ie 0;, H202, and OH·, and any organism that avails itself of the benefits of oxygen does so at the cost of maintaining an elaborate system of defenses against these intermediates We will here concern ourselves with the superoxide dismutases which, by catalytically scavenging 0;, provide a defense against it and against any reactive radical species which can be derived from it

1,139 citations

Journal ArticleDOI
TL;DR: The in-vitro development of 1-cell embryos beyond the 2-cell stage in response to the removal of glucose and the addition of glutamine to the culture medium suggests that glucose may block some essential metabolic process, and that glutamine may be a preferred energy substrate during early development for these mouse embryos.
Abstract: One-cell CF-1 x B6SJLF1/J embryos, which usually exhibit a 2-cell block to development in vitro, have been cultured to the blastocyst stage using CZB medium and a glucose washing procedure. CZB medium is a further modification of modified BMOC-2 containing an increased lactate/pyruvate ratio of 116, 1 mM-glutamine and 0.1 mM-EDTA but lacking glucose. Continuous culture of one-cell embryos in CZB medium allowed 83% of embryos to develop beyond the 2-cell stage of which 63% were morulae at 72 h of culture, but blastocysts did not develop. However, washing embryos into CZB medium containing glucose after 48 h of culture (3-4-cell stage) was sufficient to allow development to proceed, with 48% of embryos reaching the blastocyst stage by 96 h of culture. Exposure of embryos to glucose was only necessary from the 3-4-cell stage through the early morula stage since washing back into medium CZB without glucose at 72 h of culture still promoted the development of 50% of embryos to the blastocyst stage. The presence of glucose in this medium for the first 48 h of culture (1-cell to 4-cell stage) was detrimental to embryo development. Glutamine, however, exerted a beneficial effect on embryo development from the 1-cell to the 4-cell stage although its presence was not required for development to proceed during the final 48 h of culture. Blastocysts which developed under optimum conditions contained an average of 33.7 total cells. The in-vitro development of 1-cell embryos beyond the 2-cell stage in response to the removal of glucose and the addition of glutamine to the culture medium suggests that glucose may block some essential metabolic process, and that glutamine may be a preferred energy substrate during early development for these mouse embryos.

1,119 citations

Journal ArticleDOI
TL;DR: This communication describes the successful culture of one-cell to eight-cell sheep ova and one- cell and eight- cell cattle ova to the morula and blastocyst stages and reports a high embryo survival after transfer of cultured Ova to recipient animals.
Abstract: Fertilized sheep and cattle ova have not been reported to develop readily during culture in vitro. Up to 60% of sheep morulae develop normally during culture (Moor & Cragle, 1971) but earlier cleavage stages undergo limited development (Hancock, 1963; Kraemer, 1966; Tervit & McDonald, 1969; Moore, 1970) and it has been suggested that there is a block to development in vitro at the eightto twelve-cell stage (Wintenberger, Dauzier & Thibault, 1953). Only the early cleavage stages of cattle ova have been cultured and these have not been reported to develop beyond the twenty-four-cell stage in vitro (Thibault, 1966; Brinster, 1968; Sreenan, 1968; Sreenan, Scanlon & Gordon, 1968). This communication describes the successful culture of one-cell to eight-cell sheep ova and one-cell and eight-cell cattle ova to the morula and blastocyst stages and reports a high embryo survival after transfer of cultured ova to recipient animals.

1,009 citations