Development of In Vitro Matured/Fertilized Bovine Embryos in a Chemically Defined Medium: Influence of Oxygen Concentration in the Gas Atmosphere.

Takahashi, Yoshiyuki; Hishinuma, Mitsugu; Matsui, Motozumi; Tanaka, Hozumi; Kanagawa, Hiroshi

doi:10.1292/jvms.58.897

Cited by 62 publications

(67 citation statements)

References 34 publications

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“…Later studies on sheep and bovine animals [24][25][26] confirmed a positive impact of low oxygen levels on number of cells, morula and blastocyst formation. However, this improvement had no significant effect on pregnancy rates or on the viability of developing fetuses.…”

Section: Discussionmentioning

confidence: 95%

The effect of two distinct levels of oxygen concentration on embryo development in a sibling oocyte study

Kasterstein

Strassburger

Komarovsky

et al. 2013

J Assist Reprod Genet

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Purpose This prospective randomized study used sibling oocytes of 258 women with ≥8 oocytes to compare the effect of 5 % O 2 versus 20 % O 2 concentrations on embryo development and clinical outcome. Methods Oocytes of each case were divided between incubators with either 5 % or 20 % O 2 concentration. Outcome measures were fertilization, cleavage, embryo quality, blastocyst formation, and implantation, pregnancy and live birth rates. Results Fertilization and cleavage rates were similar in both groups. The 5 % O 2 group had significantly more blastomeres (P<0.05) and more top-quality embryos on day 3 (P<0.02), as well as significantly more available embryos for transfer (31.6 % vs. 23.1 % for the 20 % O 2 group; P<0.0001). There were significantly more cycles with good embryos in the 5 % group (76/258) than in the 20 % group (38/258) (P<0.0001). Implantation and pregnancy rates were significantly higher for 5 % O 2 embryos (P<0.03 and P<0.05, respectively). Live birth rates per embryo transfer were 34.2 % and 15.8 %, respectively, P<0.05. Conclusions Implantation, pregnancy and live birth rates are higher, and more good quality embryos are available for transfer and freezing with reduced rather than with atmospheric oxygen concentrations during embryo incubation.

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Section: Discussionmentioning

confidence: 95%

The effect of two distinct levels of oxygen concentration on embryo development in a sibling oocyte study

Kasterstein

Strassburger

Komarovsky

et al. 2013

J Assist Reprod Genet

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“…Oocyte classification: The cumulus-oocyte complexes (COCs) were aspirated from small antral follicles of slaughterhouse-obtained bovine ovaries as described previously [15]. They were washed two times in Hepesbuffered Tyrode's medium (TALP-Hepes) [1] supplemented with 3 mg/ml bovine serum albumin (BSA, fraction V,…”

Section: Methodsmentioning

confidence: 99%

“…They were washed two times in Hepesbuffered Tyrode's medium (TALP-Hepes) [1] supplemented with 3 mg/ml bovine serum albumin (BSA, fraction V, cumulus cells by vortexing in TALP-Hepes. They were washed three times with culture medium, a modified synthetic oviduct fluid supplemented with 20 amino acids and insulin (mSOFai) [15]. Groups of 25 to 35 presumptive zygotes were cultured in 50 µl droplets of mSOFai covered with paraffin oil at 39°C in a humidified atmosphere of 5% CO 2 , 5% O 2 and 90% N 2 .…”

Section: Methodsmentioning

confidence: 99%

“…Category 1 represented oocytes with homogeneous ooplasm and multilayered cumulus investment. Category 2 represented oocytes having heterogeneous ooplasm with dark clusters, and multilayered cumulus investment.In vitro maturation, fertilization and culture: In vitro maturation, fertilization and culture were carried out using the procedure described previously [15]. Briefly, the COCs were washed once in the maturation medium; 25 mM Hepesbuffered TCM 199 supplemented with 10% fetal calf serum (Gibco Laboratories, Grand Island, NY, U.S.A.), 0.02 units/ ml follicle-stimulating hormone (Sigma), 1 µg/ml estradiol-17β (Sigma) and 0.2 mM sodium pyruvate and 50 µg/ml gentamicin sulfate.…”

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confidence: 99%

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In Vitro Fertilization and Cortical Granule Distribution of Bovine Oocytes Having Heterogeneous Ooplasm with Dark Clusters.

Nagano

Takahashi

Katagiri

1999

The Journal of Veterinary Medical Science

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ABSTRACT. In vitro maturation, fertilization and subsequent development of oocytes with homogeneous (category 1), or heterogeneous ooplasm (category 2) were investigated. No significant differences were observed in the nuclear maturation and total fertilization rates between the two categories. However, category 2 oocytes showed a higher normal fertilization rate due to their lower incidence of polyspermy as compared to category 1 oocytes. Electron microscopic study revealed that all category 2 oocytes had cortical granules lined up next to the plasma membrane, and that some category 1 oocytes still had small clusters of cortical granules after maturation. Although the proportion of cleaved zygotes was higher in category 2, the percentages of cleaved zygotes that developed to the blastocyst stage did not differ between the two categories. These results demonstrate that oocytes with heterogeneous ooplasm have a higher capacity for normal fertilization due to the reduction in polyspermy. This can be attributed to the normal distribution of cortical granules in category 2 oocytes after maturation. -KEY WORDS: bovine, cortical granule, fertilization, morphology.J. Vet. Med. Sci. 61(5): 531-535, 1999 Sigma Chemical Co., St. Louis, U.S.A.), 0.2 mM sodium pyruvate (Sigma) and 50 µg/ml gentamicin sulfate (Sigma). During the washing procedure, oocytes were examined, selected and classified under a stereomicroscope by the following criteria. Category 1 represented oocytes with homogeneous ooplasm and multilayered cumulus investment. Category 2 represented oocytes having heterogeneous ooplasm with dark clusters, and multilayered cumulus investment.In vitro maturation, fertilization and culture: In vitro maturation, fertilization and culture were carried out using the procedure described previously [15]. Briefly, the COCs were washed once in the maturation medium; 25 mM Hepesbuffered TCM 199 supplemented with 10% fetal calf serum (Gibco Laboratories, Grand Island, NY, U.S.A.), 0.02 units/ ml follicle-stimulating hormone (Sigma), 1 µg/ml estradiol-17β (Sigma) and 0.2 mM sodium pyruvate and 50 µg/ml gentamicin sulfate. The oocytes in each category were cultured separately for 22 hr in a 50 µl droplet of maturation medium (10 or 11 oocytes per droplet) covered with paraffin oil at 39°C in a humidified atmosphere of 5% CO 2 in air. Frozen bovine semen collected from a Holstein bull was used for in vitro fertilization. After thawing the semen in a 37°C water bath for 30 sec, the motile sperm were separated using a discontinuous gradient with 45 and 90% Percoll (Pharmacia BioProcess, Uppsala, Sweden) by centrifugation at 700 × g for 20 min. The sperm were subsequently washed with a modified Brackett and Oliphant (mBO) medium [3] by centrifugation at 500 × g for 5 min. The mature oocytes were co-incubated with the sperm at the concentration of 5 × 10 6 sperm/ml in a 100 µl droplet of mBO medium containing 3 mg/ml fatty-acid-free BSA (Sigma) and 2.5 mM theophylline (Sigma) covered with paraffin oil for 20 hr at 39°C in a humidif...

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“…The culture medium was a modified synthetic oviduct fluid (mSOF) [24], with 3 mg/mL of BSA in lieu of polyvinyl alcohol. All presumptive zygotes were cultured for 150 h (7 d after starting IVF culture) as groups of 15 to 30 embryos per 50-µL droplet of mSOF covered with paraffin oil at 39 °C in a humidified atmosphere of 5% CO 2 , 5% O 2, and 90% N 2 .…”

Section: In Vitro Culture and Evaluation Of Subsequent Embryonic Devementioning

confidence: 99%

Effect of duration of in vitro maturation on nuclear maturation and fertilizability of feline oocytes

et al. 2008

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This study was conducted to improve in vitro production of embryos from domestic cats using TCM-199 as an IVM medium. The time sequence of nuclear maturation and the optimal timing of in vitro insemination were examined. Most oocytes were at the germinal vesicle stage immediately after collection; however, 8.3% had already resumed meiosis before IVM culture. After 30 h of IVM culture, the percentage of oocytes at metaphase II (MII) reached a peak (75.5%) and did not change (P>0.05) from 30 to 48 h after IVM culture. The percentage of oocytes with two pronuclei was higher (P<0.05) for oocytes matured for 30 and 36 h (38.2 and 33.0%, respectively) than for those after IVM culture for only 24 h (18.5%).Total sperm penetration rate was highest (P<0.05) for oocytes that had been matured for 30 h (46.1%). After 30 h of IVM and 18 h of IVF culture, 66.3 and 24.8% of inseminated oocytes had cleaved and developed to the blastocyst stage, respectively. We concluded that IVM of feline oocytes for 30 h in TCM-199 resulted in optimal nuclear maturation and sperm penetration.

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Development of In Vitro Matured/Fertilized Bovine Embryos in a Chemically Defined Medium: Influence of Oxygen Concentration in the Gas Atmosphere.

Cited by 62 publications

References 34 publications

The effect of two distinct levels of oxygen concentration on embryo development in a sibling oocyte study

The effect of two distinct levels of oxygen concentration on embryo development in a sibling oocyte study

In Vitro Fertilization and Cortical Granule Distribution of Bovine Oocytes Having Heterogeneous Ooplasm with Dark Clusters.

Effect of duration of in vitro maturation on nuclear maturation and fertilizability of feline oocytes

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