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中國畜牧學會會誌
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中文摘要
農科院已建立基因編輯技術進行基因剔除(Knockout, KO)，並且成功產製 KO 豬。本試驗探討 KO 公豬之新鮮精液及製成冷凍精液/解凍後之品質差異。試驗使用兩頭 KO 公豬之精液，每兩週採集 一次精液，精液冷凍以含3%甘油冷凍保護液進行，新鮮及解凍精液係以商用 BTS 稀釋後觀察， 所評估精子活力、形態完整性及頭巾完整性係以位相差顯微鏡進行。此三性狀在新鮮精液者分別為 84 ± 8%（mean ± SE; n =10）、88.6 ± 4.7% 及 79.7 ± 8%，試驗共進行 10 批次精液冷凍，並於冷 凍保存後 30 min、1 wk 及 4 wk 時，以 37℃ 水浴解凍 20 sec 後加入精液稀釋液再進行觀察評 估。試驗結果，冷凍精液於保存 30 min、1 wk 及 4 wk 後解凍，其活力分別為 53 ± 11%、46 ± 10% 及 49 ± 11%，形態完整性分別為 88.7 ± 5.9%、89.5 ± 3.7% 及 85.5 ± 5.9%，頭巾完整性分別為 65.1 ± 6.1%、62.1 ± 15.2% 及 65.5 ± 10.6%（P > 0.05）。綜合上述，KO 公豬冷凍解凍精液之精子活力、 形態完整性及頭巾完整性並未隨保存時間延長而下降，顯示此兩頭 KO 公豬之精液可進行冷凍保 存，以利於此特殊基因背景之後續保種及繁衍。
英文摘要
Gene editing technology for gene knockout (KO) has established in the Agricultural Technology Research Institute and successfully produced two lines of KO pigs. This study was conducted to evaluate the quality characteristics of frozen and thawed sperm samples obtained from KO boars. The ejaculates were collected every other week and 10 extended doses from 10 collections were used. Boar semen was frozen with 3% glycerol cryoprotectant, the fresh and thawing semen were diluted by commercial BTS extender. The sperm motility, normal morphology, and acrosome integrity were observed by phase contrast microscopy to evaluate their sperm quality. Before frozen, the motility, normal morphology and acrosome integrity of fresh semen were 84 ± 8% (mean ± SD; n =10), 88.6 ± 4.7% and 79.7 ± 8%, respectively. After frozen for 30 min, 1 wk and 4 wk, the frozen semen were thawed in 37℃ water bath for 20 sec and added into the commercial extender. After been storage for 30 min, 1 wk and 4 wk, sperm motility were 53 ± 11%, 46 ± 10%, and 49 ± 11%; normal morphology were 88.7 ± 5.9%, 89.5 ± 3.7%, and 85.5 ± 5.9%; and acrosome integrity were 65.1 ± 6.1%, 62.1 ± 15.2%, and 65.5 ± 10.6% (P>0.05), respectively. In summary, the motility, normal morphology and acrosome integrity of the frozen semen of KO boars could be maintained after thawing for at least 4 weeks of frozen storage. The results suggest that the spermatogenesis of KO boars produced are normal and their semen extenders can be treated and used as ordinary commercial boars including cryopreservation.