Development of one-cell porcine embryos to the blastocyst stage in simple media.

Hagen, D. R.; Prather, Randall S.; Sims, Michelle; First, N. L.

doi:10.2527/1991.6931147x

Cited by 85 publications

(48 citation statements)

References 7 publications

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“…Data from metabolic measurement experiments were confirmed by results of the present study and a previous study [4] where glucose supported development of in vitro produced pig embryos until the blastocyst stage. Although in vivo derived pig embryos have developed to blastocysts in the presence of glucose [16,17,24], it was recently reported that pig e m b r y o s pr o d u c e d u n d e r c u r r e n t i n v i tr o conditions have altered metabolic activity [13] that may compromise their viability. Preincubation of IVM-IVF embryos in the presence of pyruvate and lactate and subsequent culture in a glucose-supplemented medium enhanced preimplantation development, with the production of high-quality blastocysts and the birth o f p i g l e ts [ 4 ] .…”

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

“…In the pig (as in many other mammals), glucose utilization in highest in morulae and blastocysts [13,14], consistent with the energy required for the f o r m a t i o n a n d e x p a n s i o n o f b l a s t o c y s t s . Furthermore, glucose supplementation enhanced in vitro development of pig embryos [4,16,17], with the exception of the earliest preimplantation stages, where glucose appeared to have a deleterious effect on embryo development [4]. It is noteworthy that g l u c o se w as n ot c om p a ti b l e w i th i n vi tr o development of preimplantation hamster embryos [18,19] and that development of mouse embryos to the blastocyst stage was supported by pyruvate but not glucose [20].…”

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confidence: 99%

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Advanced In Vitro Production of Pig Blastocysts Obtained through Determining the Time for Glucose Supplementation

Medvedev

Ōnishi

Fuchimoto

et al. 2004

Journal of Reproduction and Development

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Abstract.The objective was to determine the effect of glucose supplementation on development (to the blastocyst stage) of in vitro matured (IVM) porcine oocytes that were either in vitro fertilized (IVF) or electrically activated (EA). Embryos were incubated for 46 or 58 h post insemination (hpi) in an NCSU37-based medium containing 0.17 mM sodium pyruvate and 2.73 mM sodium lactate (IVCPyrLac), and then transferred to an NCSU37-based medium containing 5.55 mM glucose (IVC-Glu) and cultured until Days 6 (Day 0 = day of EA or IVF). The proportions of oocytes that had formed full blastocysts by Day 6 following transfer to IVC-glu at 46 hpi was 23.5 and 41.2% in the IVF and EA groups respectively; these were lower (P<0.001) than the proportions of oocytes that formed full blastocysts after transfer at 58 hpi (60.3 and 78.7%). However, there was no significant difference in total cell number (at Day 6) between embryos transferred at 46 vs 58 hpi. We inferred that in vitroderived pig embryos can efficiently use glucose as an energy source starting at approximately 58 hpi; exposure to glucose at that time enhanced development to the blastocyst stage as well as blastocyst quality. Key words: Pig embryo, Blastocyst, Glucose usage, In vitro fertilization, Activation (J. Reprod. Dev. 50: [71][72][73][74][75][76] 2004) wine are increasingly used in transgenic technologies, including gene manipulation and nuclear transfer; both of these technologies require in vitro production of large quantities of matured oocytes and embryos. Although substantial progress has been ma de in in vitro cult ure, maturation and fertilization (IVC, IVM and IVF, respectively) of porcine oocytes, the overall efficiency of in vitro production of porcine embryos remains relatively low. Several critical aspects should be considered, i.e. variation in oocyte sources and quality of IVM [1][2][3][4][5][6], and increased polyspermy [7,8] and morphological abnormalities [4,9 ] in porcine embryos produced by IVF. Although substantial efforts have been made to optimize culture conditions for preimplantation porcine embryos, the metabolic needs of the embryo are still not well understood. In many mammals, the utilization of glucose increases as pr e im pl an t at io n d ev el op m en t p r og re s s es ; glycolysis becomes the dominant pathway during preimplantation development [10][11][12][13]. In the pig (as in many other mammals), glucose utilization in highest in morulae and blastocysts [13,14], consistent with the energy required for the f o r m a t i o n a n d e x p a n s i o n o f b l a s t o c y s t s . Furthermore, glucose supplementation enhanced in vitro development of pig embryos [4,16,17], with

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Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Advanced In Vitro Production of Pig Blastocysts Obtained through Determining the Time for Glucose Supplementation

Medvedev

Ōnishi

Fuchimoto

et al. 2004

Journal of Reproduction and Development

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“…Additional hormones, 10 IU/ml eCG (Serotropin, Teikokuzoki, Tokyo, J a p a n ) a n d 1 0 I U / m l h C G ( G o n a t r o p i n , Teikokuzoki, Tokyo, Japan), were used for oocyte maturation. The medium for oocyte collection and washing was HEPES-buffered Tyrode's medium [27] containing 0.1% (w/v) polyvinyl alcohol (TL-HEPES-PVA). The media used for IVF and sperm w a s h i n g w e r e P i g -F M a n d W S -P V A [ 2 8 ] , respectively.…”

Section: Chemicals and Mediamentioning

confidence: 99%

Effects of Delipidation and Oxygen Concentration on In Vitro Development of Porcine Embryos

Yoneda

Suzuki

Mori

et al. 2004

Journal of Reproduction and Development

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Abstract. The effects of delipidation and the oxygen (O2) concentration in the atmosphere during culture on in vitro development and H2O2 content were investigated in porcine in vivo fertilized embryos and embryos after in vitro maturation and in vitro fertilization (IVM/IVF embryos). There was no significant difference in the developmental rates to the blastocyst stage between the intact and delipidated IVM/IVF embryos. However, the mean number of cells in blastocysts derived from delipidated IVM/IVF embryos (19.8 ± 0.8 cells) was significantly smaller than that from intact embryos (24.2 ± 1.2 cells). Although there were no significant differences in the developmental rates to the blastocyst stage of intact and delipidated IVM/IVF embryos between the cultures under 5% O2 and 20% O2, the developmental rate of intact IVM/IVF embryos cultured under 5% O2 (27.1%) was significantly higher than that of the delipidated embryos cultured under 20% O2 (19.3%). On the other hand, there was no difference in the developmental rate to the blastocyst stage between in vivo fertilized embryos cultured under 5% O2 and 20% O2. Hydrogen peroxide (H2O2), one of the reactive oxygen species (ROS), is thought to cause damage to embryos. The H2O2 content per embryo derived from oocytes cultured under 5% O2 (in vivo fertilized, 58.0 ± 2.5 pixels; IVM/IVF, 79.6 ± 3.2 pixels) was significantly lower than that (in vivo fertilized, 100.2 ± 3.8 pixels; IVM/IVF, 103.9 ± 3.2 pixels) under 20% O2. Furthermore, the level of H2O2 in delipidated IVM/IVF embryos (94.7 ± 3.9 pixels) was significantly lower than that in intact embryos (103.9 ± 3.2 pixels) cultured under 20% O2. The present results indicate that the delipidation of porcine IVM/IVF embryos and reduction of the O2 concentration decreased the H2O2 level rather than the in vitro developmental rate to the blastocyst stage. Key words: Porcine embryo, In vitro development, Oxygen concentration, Delipidation, Hydrogen peroxide (J. Reprod. Dev. 50: [287][288][289][290][291][292][293][294][295] 2004) he in vitro developmental rate to the blastocyst stage of porcine in vitro maturation (IVM)/in vitro fertilization (IVF) embryos is very low compared with that of in vivo fertilized embryos [1][2][3][4]. Many obstacles must be overcome to obtain a high frequency of in vitro development in porcine I V M / I V F e m b r y o s .F o r e x a m p l e , t h e developmental competence of porcine IVM/IVF e m b r y o s i s m a rk e d l y a f f e c t e d b y t h e s a l t composition [1,5], protein supplementation [6], and small quantities of additional substances [5] in the culture medium. In addition, the oxygen (O2) concentration has been shown to be a major factor causing a difference in the developmental rates of porcine IVM/IVF embryos between in vivo and in

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“…As previously mentioned, various confounders such as concentration, pH, and ionic composition are often not accounted for. Numerous studies actually indicate HEPES is able to support oocyte maturation [45,69], fertilization [70][71][72] and embryo development [56,[72][73][74] at room atmosphere efficiently. Some studies have indicated inhibition of glucose uptake by mouse oocytes [75], lower fertilization rates [76], or compromised blastocyst formation in presence of HEPES [77], but are likely due, at least in part, to the simultaneous reductions in CO 2 levels and bicarbonate concentrations.…”

Section: Buffers In Ivfmentioning

confidence: 99%

Biological pH buffers in IVF: help or hindrance to success

Will

Clark

Swain

2011

J Assist Reprod Genet

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Purpose Minimizing environmental stress helps maintain cellular homeostasis and is a crucial component in optimizing embryo development in vitro and resulting ART success. One stressor of particular interest is pH. Biologic buffers, such as HEPES and MOPS, are valuable tools for stabilizing pH. The objective of this manuscript is to summarize efficacy and impact of various pH buffers used during IVF lab procedures Methods Keyword searches were performed using Pubmed and Medline and relevant literature reviewed. Results Various pH buffers have been used with varying degrees of success for gamete and embryo processing in a variety of animal species, as well as in human. Conclusion Though biologic buffers off a means to improve pH stability, not all buffers may be appropriate for use with gametes and embryos. Specific buffers may have undesired effects, and these may be buffer, species, cell type or concentration dependent. Continued research is needed to further refine and improve the use of biologic buffers for use in human ART.

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Development of one-cell porcine embryos to the blastocyst stage in simple media.

Cited by 85 publications

References 7 publications

Advanced In Vitro Production of Pig Blastocysts Obtained through Determining the Time for Glucose Supplementation

Advanced In Vitro Production of Pig Blastocysts Obtained through Determining the Time for Glucose Supplementation

Effects of Delipidation and Oxygen Concentration on In Vitro Development of Porcine Embryos

Biological pH buffers in IVF: help or hindrance to success

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