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題 名 | 冷凍保護液配方及平衡時間對山羊體外成熟卵母細胞玻璃化冷凍解凍後存活與後續發育能力之影響=The Viability and Subsequent Developmental Competence of In Vitro- Matured Caprine Oocytes Vitrified in Solutions with Various Cryoprotectant Formulae and Equilibrium Time |
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作 者 | 王得吉; 林信宏; 康定傑; 黃政齊; | 書刊名 | 畜產研究 |
卷 期 | 43:4 2010.12[民99.12] |
頁 次 | 頁327-337 |
分類號 | 437.58 |
關鍵詞 | 山羊; 卵母細胞; 玻璃化冷凍; 囊胚; Goat; Oocyte; Vitrification; Blastocyst; |
語 文 | 中文(Chinese) |
中文摘要 | 本計畫目的是探討不同冷凍保護劑與平衡時間對體外成熟山羊卵母細胞,經冷凍-解凍後其存活率及後續發育能力之影響。自卵巢表面直徑 2-6 mm 之濾泡中取得卵丘卵母細胞複合體,隨即進行體外成熟作用。在玻璃化冷凍前,體外成熟卵母細胞利用二階段平衡方式,分別以 7.5% EG (ethylene glycol)+ 7.5% DMSO (dimethyl sulfoxide)7 分鐘、10% EG + 10% DMSO 45 秒及 1.6M EG 6 分鐘與 12 分鐘進行第一階段平衡後,再分別以 16.5% EG + 16.5% DMSO 25 秒、20% EG+ 20% DMSO 25 秒及 6.8 M EG 30 秒進行第二階段短暫平衡後,於玻璃細管尖端形成之每一微滴含 5 個卵母細胞的方式,直接投入液態氮中進行玻璃化冷凍與長期保存。解凍後卵母細胞分別觀察其恢復正常外觀比率及經體外受精後體外培養至囊胚之比率,以評估其存活率及後續發育能力。試驗結果顯示,體外成熟後之卵母細胞利用 7.5% EG + 7.5% DMSO 之冷凍保護劑預先平衡 7 分鐘,再轉移至 16.5% EG + 16.5% DMSO 冷凍保護劑平衡 25 秒後進行微滴玻璃化冷凍保存,其解凍後形態恢復比例顯著 (P < 0.05) 高於其他處理組,而在後續發育能力亦有較佳之趨勢。 |
英文摘要 | The objective of this study was to examine the effects of cryoprotectants and equilibrium time on the survival rate and the developmental potential of vitrified-warmed caprine oocytes matured in vitro. Cumulusoocyte complexes (COCs) obtained from 2-6 mm follicles on the surface of ovaries were cultured in vitro for maturation. Before vitrification, two-step of equilibration of in vitro matured oocytes in solutions containing different concentration of cryoprotectants for different time were adopted, i. e., 7.5% EG+7.5% DMSO for 7 min, 10% EG+10% DMSO for 45 sec and 1.6 M EG for 6 min or 12 min and then to 16.5% EG+16.5% DMSO for 25 sec, 20% EG+20% DMSO for 25 sec and 6.8 M EG for 30 sec. A group of five oocytes in a 2 μ l microdrop of the vitrification solution generated from the tip of a glass pipette were eventually flipped into liquid nitrogen for vitrification and long-term cryopreservation. The survival rate and developmental competence of vitrified-warmed oocytes were evaluated by the morphology of oocytes after warming and the blastocyst formation rate following fertilize and culture in vitro. The results indicated that in vitro matured oocytes equilibrated in solution containing 7.5% EG+7.5% DMSO for 7 min and then transferred to 16.5% EG+16.5% DMSO for 25 sec resulted in the highest survival rate (P < 0.05) and tended to have better developmental competence to blastocyst than that in the rest of treatments. |
本系統中英文摘要資訊取自各篇刊載內容。