体外受精由来牛胚盤胞期胚の染色体検査における培地へのヘパリン添加の影響

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The optimum concentration of heparin for obtaining high incidence of analysable metaphase spreads was investigated using early, expanded and hatched bovine blastocysts derived from in vitro fertilization.The ebryos were treated with TCM 199 containing colcemid (0.04μg/ml) either with heparn (100, 200 or 400μg/ml) or without heparin at 39° for 2hr before they were subjected to a short-term treatment with 0.5% sodium citrate solution. The addition of 100 or 200μg/ml heparin resulted in higher incidences of metaphases and higher mitotic indexes. A dose of 200μg/ml heparin resulted in the highest incidences of metaphases and mitotic indexes in expanded and hatched blastocysts, mitotic indexes being 6.2, 7.2 and 4.4% for early, expanded and hatched blastocysts, respectively. The rates of embryos with metaphase plates in the treatment of 200μg/ml heparin were 94.4 (51/54), 96.6 (56/58) and 100% (31/31) for early, expanded and hatched blastocysts, respectively. The sexing rates in the treatment of 200μg/ml heparin were the highest in each developmental stages; 72.2 (39/54), 91.4 (53/58) and 90.3% (28/31) for early, expanded and hatched blastocysts, respectively. By contrast, the sexing rates in the absence of heparin were 43.8 (21/48), 60.7 (34/56) and 57.7% (15/26), respectively. The addition of 400μg/ml heparin did not increase the incidence of metaphases and mitotic index and did not increase the sexing rate. These results suggest that 200μg/ml heparin promotes the viability and mitotic activity of bovine blastocysts derived from in vitro fertilization during the colcemid-treatment.
社団法人日本畜産学会の論文

体外受精由来牛胚盤胞期胚の染色体検査における培地へのヘパリン添加の影響

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