マウス初期胚の染色体検索に関する研究

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  • A methodological study on chromosomal preparation of mouse embryos.
  • マウス ショキ ハイ ノ センショクタイ ケンサク ニ カンスル ケンキュウ

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A total of 304 mouse expanded blastocysts were cultured in a CO2 incubator at 37 C to obtain the optimum conditions, namely, the kind of medium, the concentration of colchicine and duration of culture for chromosomal analysis. Criteria used to determine the optimum culture conditions were the number of nuclei, metaphase plates and size of chromosomes. In the culture by BMOC-3, it was significant that the largest number of metaphases was observed compared with the culture by MEM+ 20%FCS and PBS+20%FCS (P<0.05). In the culture of 0.4-1.6μg/ml colchicine in the BMOC-3, more metaphases were obtained than that of 0.025-0.05μg/ml colchicine. The number of metaphases increased with the extension of culture period up to 12 h, but the number of nuclei decreased con-versely because of the degeneration of cells. The contraction of chromosomes was observed in the culture of over 6 h with colchicine. From the results, it was concluded that the 2-hour culture by BMOC-3 containing 0.4μg/ml colchicine is the optimum condition for chromosomal preparation in this study.<BR>Under this condition, a total of 155 expanded blastocysts from 3 weeks young females of ddY strain were examined. They had an average of 66.0 nuclei and 5.7 metaphases per embryo. The 145 blastocysts out of 155 blastocysts showed explicitly countable metaphases, and normal diploidy was found in 118 blastocysts (81.8%). Out of 529 metaphases from 145 blastocysts, only 272 metaphases (51.4%) showed a normal diploidy, while 156 metaphases (29.5%) showed hypodiploidy. It seems that a slight loss of chromosomes from metaphase plate is inevitable in the chromosomal analysis of em-bryos.

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