Time filter

Source Type

Dang-Nguyen T.Q.,Japan National Agriculture and Food Research Organization | Dang-Nguyen T.Q.,Veterinary Research Institute | Haraguchi S.,Japan National Agriculture and Food Research Organization | Kikuchi K.,Japan National Institute of Agrobiological Science | And 5 more authors.
Molecular Reproduction and Development | Year: 2014

SUMMARY: We produced recombinant porcine leukemia inhibitory factor (pLIF) and examined its effect on in vitro maturation (IVM) of porcine oocytes and their developmental competence after in vitro fertilization. Porcine cumulus-oocyte complexes (COCs) were matured in a medium supplemented with pLIF during the first 22hr, last 22hr, or entire 44hr duration of IVM. Oocytes in all groups tended to show enhanced nuclear maturation rates by the metaphase II (MII) stage (76.1%, 82.1%, and 86.6%, respectively) compared to the without-pLIF treatment group (69.6%, control). A significant increase in MII rate (P<0.05) and obvious induction of cumulus expansion were observed over the whole time span (44hr) in the IVM group. When cumulus cells were removed at 22hr and denuded oocytes were further cultured, pLIF showed no effect on maturation rate. Oocytes matured in pLIF-supplemented medium showed a tendency for more rapid blastocyst development (21.1% vs. 16.2%, P=0.0715). Examination of transcripts and proteins of the LIF signaling pathway in COCs revealed that LIF, LIF receptors, and signal transducer and activator of transcription 3 (STAT3) are present in both cumulus cells and oocytes. The amount of phosphorylated STAT3 (p-STAT3) markedly increased in both cumulus cells and oocytes cultured in pLIF-supplemented media, although oocyte p-STAT3 disappeared after 44hr of IVM. These results suggest that the LIF/STAT3 pathway is functional during IVM of porcine oocytes, and supplementing pLIF in the IVM medium can improve oocyte maturation by activating this pathway. © 2013 Wiley Periodicals, Inc.


Chasombat J.,Khon Kaen University | Nagai T.,Food and Fertilizer Technology Center | Nagai T.,Japan National Agriculture and Food Research Organization | Parnpai R.,Suranaree University of Technology | Vongpralub T.,Khon Kaen University
Cryobiology | Year: 2015

The stabilization of spindle fibersis important for successful vitrification of bovine oocytes because microtubules and other cytoskeleton fibers (CSF) can be damaged during vitrification, resulting in failure of fertilization after thawing. Docetaxel, a stabilizing agent, could potentially reduce CSF damage of bovine oocytes induced during vitrification. However, there have been no reports on the effects of docetaxel on their vitrification. Experiment 1 was conducted to investigate the effects of various doses of docetaxel (0.0, 0.05, 0.5, 5.0 and 50. μM) in preincubation medium of in vitro matured (IVM) bovine oocytes on their developmental ability after in vitro fertilization (IVF). The results show that 0.05. μM docetaxel had no adverse effect on embryo development, while docetaxel at a concentration of ≥0.5. μM inhibited development. Experiments 2 and 3 were conducted to investigate the effects of preincubation of IVM bovine oocytes with 0.05. μM docetaxel for 30. min prior to vitrification-warming on CSF integrity (Experiment 2), and on oocyte survival and viability after IVF (Experiment 3). When preincubated with 0.05. μM docetaxel for 30. min before vitrification, post-thawed oocytes had less CSF damage and higher survival rates compared with those untreated with docetaxel before vitrification. Surviving oocytes also had higher rates of cleavage and development to the blastocyst stage after IVF. In conclusion, preincubation of IVM bovine oocytes with 0.05. μM docetaxel for 30. min before vitrification was effective at preventing CSF damage during vitrification, and improving oocyte viability after warming and subsequent cleavage and blastocyst formation after IVF. © 2015 Elsevier Inc.


Suttirojpattana T.,Japan National Agriculture and Food Research Organization | Suttirojpattana T.,Suranaree University of Technology | Somfai T.,Japan National Agriculture and Food Research Organization | Matoba S.,Japan National Agriculture and Food Research Organization | And 4 more authors.
Theriogenology | Year: 2016

The aim of the present study was to optimize the temperature for the temporal storage of matured bovine oocytes. In vitro-matured bovine oocytes were preserved in HEPES-buffered TCM199 medium supplemented with 10% newborn calf serum at different temperatures (4 °C, 15 °C, 25 °C, and 38.5 °C) for 20 hours. Embryo development and blastocyst quality after in vitro fertilization, cytoplasmic ATP and glutathione levels in oocytes, and the frequency of apoptotic oocytes were compared among storage groups and a control group without storage. Among the storage groups, those at 25 °C and 38.5 °C showed the highest rates of blastocyst development (19.3% and 24.5%, respectively) compared with those stored at 4 °C and 15 °C (8.5% and 14.9%, respectively); however, blastocyst formation rates in all storage groups were lower than that in the control group (39.8%; P < 0.05). Storage at 38.5 °C and 15 °C was associated with reduced cell numbers in resultant blastocysts compared with the control and the 25 °C storage groups. Storage at 4 °C reduced metabolic activity of oocytes characterized by their lower ATP levels compared with the other groups. Storage for 20 hours significantly reduced the glutathione content in oocytes in all groups in a similar manner, irrespective of the temperature. Storage at 4 °C or 15 °C but not at 25 °C and 38.5 °C significantly increased the percentage of apoptotic oocytes compared with the control group. In conclusion, 25 °C was found to be the most suitable temperature for the temporal storage of matured bovine oocytes regarding both the developmental competence of oocytes and the quality of resultant blastocysts. © 2016 Elsevier Inc.


Parnpai R.,Suranaree University of Technology | Liang Y.,Suranaree University of Technology | Ketudat-Cairns M.,Suranaree University of Technology | Somfai T.,Japan National Agriculture and Food Research Organization | And 3 more authors.
Theriogenology | Year: 2016

During the past decade, vitrification has been acknowledged as an efficient alternative to traditional slow-rate freezing in both human and animal embryology. The buffalo is the major milk and meat producing farm animal in many developing countries. Cryopreservation of buffalo oocytes and embryos is very important in preserving this species for future use. This review discusses the recent buffalo oocytes and embryos vitrification procedures, different types of cryoinjuries, and other factors affecting the vitrification of buffalo oocytes and embryos. © 2016 Elsevier Inc.


Takahashi T.,Akita | Sasaki K.,Akita | Somfai T.,Japan National Agriculture and Food Research Organization | Nagai T.,Food and Fertilizer Technology Center | And 3 more authors.
Journal of Reproduction and Development | Year: 2016

The antioxidant effect of N, N-dimethylglycine (DMG) on in vitro-produced (IVP) bovine embryos was examined. After in vitro fertilization, presumptive zygotes were cultured with or without 0.1 µM DMG under different oxygen tensions. The percentage of embryos developing to the blastocyst stage was lowest under a 20% oxygen concentration without DMG, and it was significantly increased (P < 0.05) by applying a 5% oxygen concentration. Under the 20% oxygen concentration, supplementation of the medium with DMG significantly improved blastocyst development, which was nearly equal to that achieved under 5% oxygen without DMG. Furthermore, a tendentious increase (P = 0.06) in blastocyst cell numbers was observed when DMG was applied. In the second experiment, addition of H2O2 (0.5 mM) to the culture medium significantly (P < 0.01) reduced the percentage of embryos developing to the blastocyst stage. However, DMG supplementation prevented this reduction. In conclusion, DMG enhanced the in vitro development of IVP bovine embryos by acting as an antioxidant. © 2016 by the Society for Reproduction and Development.


Inaba Y.,Japan National Agriculture and Food Research Organization | Inaba Y.,National Livestock Breeding Center Tokachi Station | Abe R.,Miyagi Prefecture Tobu Livestock Hygiene Service CenterMiyagi | Geshi M.,Japan National Agriculture and Food Research Organization | And 4 more authors.
Journal of Reproduction and Development | Year: 2016

The aim of the present study was to clarify if flow-cytometric sex-sorting of bovine sperm affected in vitro blastocyst production in different bulls, either in terms of its ability to fertilize the oocyte or by interfering with post-fertilization embryo development. We performed in vitro fertilization (IVF) using both commercially available frozen-thawed X-sorted and non-sorted sperm of 4 Holstein bulls at 3 concentrations (1 × 106, 2 × 106, and 5 × 106 sperm/ml). When fertilization rates were compared, a variation in fertilization rates among different sperm concentrations was detected in 2 bulls, with similar results for X-sorted and non-sorted sperm. However, we found no evidence that the fertilization rates were affected by the sorting process. To investigate effects on embryo development, we determined the optimum sperm concentration for IVF in each bull, which resulted in similar fertilization rates among bulls. We next performed IVF using both X-sorted and non-sorted sperm of the 4 bulls at their optimum sperm concentration and compared in vitro embryo development. Cleavage rates with X-sorted sperm were similar to their non-sorted counterparts. However, significantly reduced blastocyst development was associated with the use of X-sorted sperm in one bull, whereas in the other three bulls, blastocyst development after IVF with X-sorted and non-sorted sperm was similar. In conclusion, in our system, X-sorting affects in vitro blastocyst production by reducing the developmental competence of fertilized oocytes rather than affecting the fertilization ability of the sperm. However, the occurrence of this phenomenon varies among bulls. © 2016 by the Society for Reproduction and Development.


Somfai T.,Japan National Agriculture and Food Research Organization | Yoshioka K.,Japanese National Institute of Animal Health | Tanihara F.,Japan National Institute of Agrobiological Science | Tanihara F.,Yamaguchi University | And 6 more authors.
PLoS ONE | Year: 2014

We report the successful piglet production from cryopreserved oocytes for the first time by using a simple, high capacity vitrification protocol for preservation and a defined system for in vitro embryo production. Immature cumulus-oocyte complexes (COCs) from prepubertal gilts were vitrified in microdrops and stored in liquid nitrogen. After warming, COCs were subjected to in vitro maturation (IVM), fertilization (IVF), and subsequent culture (IVC). Adjusting warmplate temperature to 42°C during warming prevented temperature drops in a medium below 34.0°C and significantly increased the percentage of oocyte survival and thus blastocyst yields obtained from total vitrified oocytes compared with that of warming at 38°C (87.1% vs 66.9% and 4.4% vs 2.7%, respectively). Nuclear maturation and fertilization of oocytes were not affected by vitrification and warming temperature. Blastocyst development on day 7 (day 0 = IVF) of the surviving oocytes after warming at 38°C and 42°C was not different but lower (P<0.05) than those of non-vitrified control oocytes (4.6%, 5.2% and 17.9%, respectively). However, blastocyst cell numbers in the control and vitrified groups were similar irrespective of warming temperature. Omitting porcine follicular fluid (pFF) from IVM medium (POM) did not affect maturation, fertilization and embryo development of vitrified-warmed oocytes. Transfer of blastocysts obtained on day 5 from vitrified oocytes matured either with or without pFF into 4 recipients (2 for each group) resulted in 4 pregnancies and the delivery of a total of 18 piglets. In conclusion, optimization of warming temperature was a key factor for achieving high survival rates, and surviving oocytes could be utilized in vitro using defined media. Using these modifications, live piglets could be obtained from cryopreserved oocytes for the first time. © 2014 Somfai et al.


PubMed | Suranaree University of Technology, Japan National Agriculture and Food Research Organization and Food and Fertilizer Technology Center
Type: | Journal: Animal science journal = Nihon chikusan Gakkaiho | Year: 2016

Our aim was to improve the developmental competence of bovine oocytes during their liquid storage by using additives. In vitro matured oocytes were stored for 20h at 25C in HEPES buffered TCM 199 medium (base medium). After storage, in vitro embryo development after in vitro fertilization was compared to those of non-stored (control) ones. Addition of 10% (v/v) newborn calf serum or 10.27mmol/L pyruvate alone to the base medium did not improve blastocyst formation rates in stored oocytes; however, their simultaneous addition significantly improved the rate compared with those stored in base medium (P<0.05). Supplementation of the holding medium with dithiothreitol (DTT) at any concentrations did not improve embryo development from stored oocytes. Although supplementation with cyclosporine A (CsA) significantly reduced apoptosis and membrane damage rates during storage, it did not improve the developmental competence of oocytes. 1,2-bis(2-aminophenoxy) ethane N,N,N,N-tetraacetic acid tetrakis-acetoxymethyl ester and ruthenium red had no effect on oocyte apoptotic rates. Blastocyst formation rates in all stored groups remained significantly lower than that of the control. In conclusion, pyruvate and serum had a synergic effect to moderate the reduction of oocyte quality during storage, whereas mitochondrial membrane pore inhibitor CsA and the antioxidant DTT did not affect their developmental competence.


PubMed | Japan National Institute of Agrobiological Science, National Livestock Breeding Center, Japan National Agriculture and Food Research Organization and Food and Fertilizer Technology Center
Type: Journal Article | Journal: Animal science journal = Nihon chikusan Gakkaiho | Year: 2015

Follicle stimulation by follicular stimulating hormone (FSH) is known to improve developmental competence of bovine oocytes obtained by Ovum Pick-Up (OPU); however, the exact factors in oocytes affected by this treatment have remained unclear. We compared in vitro matured (IVM) oocytes obtained at the immature stage from cows by OPU either without or with stimulation with FSH (non-stimulated and stimulated OPU, respectively) to those obtained by superstimulation and in vivo maturation in terms of cytoskeleton morphology, mitochondrial distribution, intracellular adenosine triphosphate (ATP) content and H2 O2 levels at the metaphase-II stage and intracellular Ca(2+) levels after in vitro fertilization (IVF). Confocal microscopy after immunostaining revealed reduced size of the meiotic spindle, associated with increased tendencies of microfilament degradation and insufficient mitochondrial re-distribution in non-stimulated OPU-derived IVM oocytes compared with those collected by stimulated OPU, which in turn resembled in vivo matured oocytes. However, there was no difference in mitochondrial functions between oocytes obtained by stimulated or non-stimulated OPU in terms of ATP content, cytoplasmic H2 O2 levels, base Ca(2+) levels and the frequencies and amplitudes of Ca(2+) oscillations after IVF. Larger size of metaphase spindles in oocytes obtained by stimulated OPU may reflect and potentially contribute to their high developmental competence.


Lee C.-F.,Agricultural Research Institute | Chang H.-Y.,Tainan District Agricultural Research and Extension Station | Wang C.-L.,Agricultural Research Institute | Chen W.-S.,Food and Fertilizer Technology Center
Zoological Studies | Year: 2011

A review of Phyllotreta Chevrolat in Taiwan (Coleoptera: Chrysomelidae: Galerucinae: Alticini). Zoological Studies 50(4): 525-533. Species of the genus Phyllotreta Chevrolat in Japan and Taiwan are reviewed. Phyllotreta striolata (Fabricius) is redescribed based on Taiwanese specimens; P. chotanica Duvivier is recorded for the 1st time in Taiwan; and P. downesi insularis Heikertinger is raised to species level. Lectotypes and paratypes are designated for P. downesi insularis.

Loading Food and Fertilizer Technology Center collaborators
Loading Food and Fertilizer Technology Center collaborators