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Atsushi IDETA Yoshito AOYAGI Kanami TSUCHIYA Yuuki NAKAMURA Kou HAYAMA Atsushi SHIRASAWA Kenichiro SAKAGUCHI Naomi TOMINAGA Yoshiyuki NISHIMIYA Sakae TSUDA 《The Journal of reproduction and development》2015,61(1):1-6
Embryos obtained via superovulation are necessary for mammalian artificial reproduction, and viability is a key determinant of success. Nonfreezing storage at 4 C is possible, but currently used storage solutions can maintain embryo viability for only 24–48 h. Here we found that 10 mg/ml antifreeze protein (AFP) dissolved in culture medium 199 with 20% (v/v) fetal bovine serum and 25 mM HEPES could keep bovine embryos alive for 10 days at 4 C. We used a recombinant AFP isolated from the notched-fin eelpout (Zoarces elongatus Kner). Photomicroscopy indicated that the AFP–embryo interaction was enhanced at 37 C. Embryos pre-warmed with the AFP solution at 37 C for 60 min maintained high viability, whereas those that were not pre-warmed could live no longer than 7 days. Thus, short-term storage of bovine embryos was achieved by a combination of AFP-containing medium and controlled pre-warming. 相似文献
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Chie SUZUKI Yosuke SAKAGUCHI Hiroyoshi HOSHI Koji YOSHIOKA 《The Journal of reproduction and development》2016,62(1):79-86
The effects of lipid-rich bovine serum albumin (LR-BSA) on the development of porcine blastocysts produced
in vitro were examined. Addition of 0.5 to 5 mg/ml LR-BSA to porcine blastocyst medium
(PBM) from Day 5 (Day 0 = in vitro fertilization) significantly increased the hatching rates
of blastocysts on Day 7 and the total cell numbers in Day-7 blastocysts. When Day-5 blastocysts were cultured
with PBM alone, PBM containing LR-BSA, recombinant human serum albumin or fatty acid-free BSA, addition of
LR-BSA significantly enhanced hatching rates and the cell number in blastocysts that survived compared with
other treatments. The diameter, ATP content and numbers of both inner cell mass and total cells in Day-6 and
Day-7 blastocysts cultured with PBM containing LR-BSA were significantly higher than in blastocysts cultured
with PBM alone, whereas LR-BSA had no effect on mitochondrial membrane potential. The mRNA levels of enzymes
involved in fatty acid metabolism and β-oxidation (ACSL1, ACSL3,
CPT1, CPT2 and KAT) in Day-7 blastocysts were
significantly upregulated by the addition of LR-BSA. The results indicated that LR-BSA enhanced hatching
ability and quality of porcine blastocysts produced in vitro, as determined by ATP content,
blastocyst diameter and expression levels of the specific genes, suggesting that the stimulatory effects of
LR-BSA arise from lipids bound to albumin. 相似文献
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Several lines of evidence implicate small spirochetes, presumably treponemata, as etiologic agents in the production of the maternally transmitted "sex ratio" condition (SR) in Drosophila nebulosa, in D. willistoni, and in strains of D. melanogaster into which the SR condition has been artificially transferred. The presence of treponemata in the hemolymph of adult females of these species is completely correlated with the production of unisexual progenies and like this condition is dependent on the genotype of the host and of the infectious agent. 相似文献
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Kenichiro Sakaguchi Excel Rio S. Maylem Ramesh C. Tilwani Yojiro Yanagawa Seiji Katagiri Edwin C. Atabay Eufrocina P. Atabay Masashi Nagano 《Animal Science Journal》2019,90(5):690-695
In this study, we examined the effects of superstimulation using follicle‐stimulating hormone (FSH) followed by gonadotropin‐releasing hormone (GnRH) on buffalo embryo production by ultrasound‐guided ovum pick‐up (OPU) and in vitro fertilization (IVF). Nine Murrah buffaloes were subjected to OPU‐IVF without superstimulation (control). The morphologies of the oocytes collected were evaluated, and oocytes were then submitted to in vitro maturation (IVM). Two days after OPU, same nine buffaloes were treated with twice‐daily injections of FSH for 3 days for superstimulation followed by a GnRH injection. Oocytes were collected by OPU 23–24 hr after the GnRH injection and submitted to IVM (the superstimulated group). The total number of follicles, number of follicles with a diameter > 8 mm, and number of oocytes surrounded by multi‐layered cumulus cells were higher in the superstimulated group than in the control group (p ≤ 0.05). After IVF, the percentages of cleavage and development to blastocysts were higher in the superstimulated group than in the control group (p < 0.05). In conclusion, superstimulation improved the quality of oocytes and the embryo productivity of OPU‐IVF in river buffaloes. 相似文献
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Tamura K Arai H Ueno E Saito C Yagihara H Isotani M Ono K Washizu T Bonkobara M 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2007,69(9):925-930
Dendritic cell (DC) vaccination is one of the most attractive immunotherapies for malignancies in dogs. To examine the differences in DC-mediated immune responses from different types of malignancies in dogs, we vaccinated dogs using autologous DCs pulsed with keyhole limpet hemocyanin (KLH) and cell lysate prepared from squamous cell carcinoma SCC2/88 (SCC-KLH-DC), histiocytic sarcoma CHS-5 (CHS-KLH-DC), or B cell leukemia GL-1 (GL-KLH-DC) in vitro. In vivo inductions of immune responses against these tumor cells were compared by the delayed-type hypersensitivity (DTH) skin test. The DTH response against SCC2/88 cells were observed in dogs vaccinated with autologous SCC-KLH-DC, while the response was undetectable against CHS-5 and GL-1 cells in dogs vaccinated with autologous CHS-KLH-DC and GL-KLH-DC. Skin biopsies taken from DTH challenge sites were then examined for immunohistochemistry, and recruitment of CD8 and CD4 T cells was detected at the site where SCC2/88 cells were inoculated in dogs vaccinated with SCC-KLH-DC. By contrast, neither CD8 nor CD4 T cell infiltration was found at the DTH challenge site in the dogs vaccinated with CHS-KLH-DC or GL-KLH-DC. These findings may reflect that the efficacy of immune induction by DC vaccination varies among tumor types and that immune responses could be inducible in squamous cell carcinoma. Our results encouraged further investigation of therapeutic vaccination for dogs with advanced squamous cell carcinoma in clinical trials. 相似文献
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Uematsu Y Yamaguchi T Koike A Yagihara H Hasegawa D Matsuki N Ono K Washizu T Arai T Bonkobara M 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2008,70(8):845-847
A monoclonal antibody, K9BYU, was generated using Escherichia coli recombinant extracellular domain of canine neural-cell adhesion molecule (N-CAM) as an antigen. Immunoreactivity of K9BYU to insect cell recombinant canine N-CAM was demonstrated by Western blotting using Sf9 insect cells transfected with the canine N-CAM gene. In Western blotting against canine brain tissue, K9BYU detected three isoforms of N-CAM that correspond to three major isoforms of human and mouse N-CAM (N-CAM-120, -140, and -180). From these results, K9BYU was considered to be a useful tool for research of canine N-CAM. 相似文献
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