@article{oai:az.repo.nii.ac.jp:00005144,
 author = {紫野, 正雄 and 柏崎, 直巳 and 猪股, 智夫 and Shino, Masao and Kashiwazaki, Naomi and Inomata, Tomo},
 journal = {麻布大学雑誌, Journal of Azabu University},
 month = {Mar},
 note = {ブタ体細胞核移植胚の発生能は低く,産子作出効率は極めて低い。しかし,体細胞核移植を介したトランスジェニックブタの作製法は,DNA顕微注入法よりも有利な点が多い。本研究はブタ体細胞核移植胚の発生能の改善を目的とし,単為発生胚と体細胞核移植胚を集合させ,その集合胚の発生能を検討した。実験1: ブタ初期胚の集合胚作製条件を検討するために,発生ステージが異なる単為発生胚どうしの集合胚の集合率および胚盤胞への発生率を調べた。単為発生胚は,48h体外成熟培養後の卵母細胞に1kV/cm,64μsec,0.5sec間隔で三回印可による活性化誘起処理を施し,サイトカラシンBで倍数体処理をおこなって作出した。活性化処理後48あるいは72hに05%pronaseにより透明帯を除去し,発生ステージの異なる単為発生胚を3通りの組み合わせ(48h+48h区,48h+72h区,72h+72h区)で集合させ,その集合率,胚盤胞への発生率および胚盤胞の細胞数を調べた。活性化処理48h後に透明帯除去し,集合させずに培養したものを対照区とした。透明帯除去単為発生胚の集合率および胚盤胞への発生率は,48h+48h区で58.3%(60/103)および50.5%(52/103),48h+72h区で28.5%(43/151)および22.5%(34/151),72h+72h区で38.6%(71/184)および28.8%(53/184)となり,48h+48h区が他の区に比べ有意に(P<0.05)高い値を示した。また,対照区は胚盤胞への発生率が36.7%(18/49)で,48h+48h区より有意に(P<0.05)低い値を示した。集合胚由来の胚盤胞における細胞数は,各区が対照区に比べ有意に(P<0.05)高い値を示した。実験2: 実験1の結果から,活性化処理後48hに透明帯除去させた単為発生胚と卵丘細胞をドナーとした核移植胚とを集合させ(集合区),その集合率,胚盤胞への発生率および胚盤胞の細胞数を調べた。活性化処理48h後に透明帯除去し,集合させなかった核移植胚を対照区とした。集合胚由来の胚盤胞への発生率および胚盤胞の細胞数は38.2%(39/102)および55.3±4.3,対照区では20.9%(9/43)および25.3±2.5であり,集合区が対照区に比べ有意に(P<0.05)高い胚盤胞への発生率を示した。以上の結果から,ブタ体細胞核移植胚と単為発生胚をともに活性化処理後48hに集合させることで,ブタ体細胞核移植胚の胚盤胞への発生能が改善することが示唆された。, Somatic cell nuclear transfer cloned (SCNT) embryos have still lower developmental competence in the pig. However, transgenesis through somatic cell cloning in pigs has more advantage than conventional DNA micro-injection method. The aim of the present study was to improve the competence of SCNT porcine embryos, we examined that embryonic development in vitro of aggregated embryos between SCNT and parthenogenetic embryos. In Experiment I, rates of successful aggregation and development to the blastocyst stage were assessed. In vitro matured porcine oocytes were electro-stimulated (1kV/cm, 64μsec, three times, interval of 0.5sec) and cultured with 5μg cytochalasin B for 2 hours (h) to obtain diploid parthenogenetic embryos. At 48 or 72h, the zona pellucidae of activated oocytes were removed with 0.5% pronase, the zona-free embryos were treated with aggregation (48h+48h, 48h+72h, 72h+72h), and cultured in vitro. The zona-free embryos at 48h post electro-activation were also cultured as control. The successful aggregation rates and the blastocyst formation rates were 58.6% (60/103) and 50.5% (52/103) in the 48h+48h treatment, 28.5% (43/151) and 22.5% (34/151) in the 48h+72h tratment, and 38.6% (71/184) and 28.8% (53/184) in the 72h+72h treatment, respectively. The rate of the 48h+48h treatment was significantly (P<0.05) higher than other treatment groups. In addition, the blastocyst formation rate of the 48h+48h treatment was significantly (P<0.05) higher than the control (36.7%, 18/49). In Experiment II, the SCNT embryos and parthenogentic embryos at 48h post-activation were treated with aggregation, and cultured in vitro. The successful aggregation rate and the blastocyst formation rate were 38.2% (39/102) and 55.3±4.3%, respectively, and the blastocyst formation rate was significantly (P<0.05) higher than the control (20.9%, 9/43). The results of the present study indicated that development to the blastocysts stage of porcine SCNT embryos aggregated with parthenogenetic embryos was improved., P(論文), 特集, application/pdf, FEATURE ARTICLES},
 pages = {193--199},
 title = {ブタ体細胞核移植胚の発生能改善に関する研究},
 volume = {13/14},
 year = {2007},
 yomi = {シノ, マサオ and カシワザキ, ナオミ and イノマタ, トモオ}
}