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題名 | 源自不同生長期別豬腔前濾泡之卵母細胞於離體之生長和後續之成熟率=Effect of Porcine Preantral Follicle at Different Stages of Development on Growth and Subsequent Maturation of Oocyte in Vitro |
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作者 | 鄭三寶; 顏大慶; 顏淑卿; 劉炳燦; 邱啟明; Cheng, San-pao; Yen, Da-ching; Yen, Su-ching; Liu, Bing-tsan; Chiou, Chii-ming; |
期刊 | 中華農學會報 |
出版日期 | 19970600 |
卷期 | 178 1997.06[民86.06] |
頁次 | 頁104-119 |
分類號 | 437.21 |
語文 | chi |
關鍵詞 | 腔前濾泡; 卵母細胞; 存活率; 成熟率; 離體生長; 豬; Preantral follicle; Oocyte; Survival rate; Maturation rate; Growth in vitro; Porcine; |
中文摘要 | 本試驗旨在評估不同生長期別之腔前濾泡于離體發育過程,其內卵母細胞之直徑 和存活率以及後續成熟率之變化。 腔前濾泡內卵母細胞之存活率, 係應用 FDA (3',6'-diacetyl fluorescin ) 螢光染色技術予以判定。 依培養前腔前濾泡內卵母細胞之 直徑所區分為 <7O μ m (小型 )、70 ~<8O μ m (中型 ) 和 80 ~ 9O μ m (大型 ) 等三 組不同生長期別之腔前濾泡,經海藻酸鈣膠層包埋後,置于生長培養液中培養 16 天之平均 直徑分別自培養前之 260、320 和 390um 增加至培養後之 520、600 和 610 μ m,卵母細 胞之平均直徑由 58、 71 和 84 μ m 增加至 86、106 和 114 μ m; 而腔狀濾泡之形成率 分別為 0.4 和 14%。 惟三組卵母細胞之存活率, 分別自培養後第 4 天之 39、53 和 63% 下降至第 16 天之 10、28 和 25%。大型卵母細胞組之濾泡和卵母細胞之直徑以及卵母細胞 之存活率等,於培養期間均持續高於小型組者。 當比較於離體生長 (由直徑 80 ~ 9O μ m 卵母細胞之腔前濾泡經離體培養 12 天者 ),和於體內生長 [ 取自卵巢之腔狀濾泡 ( 直徑 >l mm) 者 ] 之卵丘卵母細胞複合體,經離體成熟培養 44 ~ 48 hr 之結果顯示,不論於體 內或離體所生長之小型卵母細胞 (直徑 <9O μ m),均未能在離體成熟培養中恢復減數分裂 ,以迄生發泡瓦解 (GVBD) 階段。就直徑 >9O μ m 卵母細胞而言,於體內所生長之卵母細 胞之成熟率極顯著高於離體所生長者 [61% (169/274) vs . 2% (2/103),P<O.01] 。綜合 前述資料提示,腔前濾泡於離體培養發育和卵母細胞之存活率等,均與培養前該濾泡之卵母 細胞之直徑關係密切; 於體內所生長之腔狀濾泡之卵母細胞,其成熟率隨其直徑之增加而提 高。 |
英文摘要 | Effect of isolated preantral follicle at different stages of development on growth and subsequent maturation of porcine oocytes in vitro was conducted in this study. Ovaries were collected from prepubertal gilts at a local abattoir. Minced ovaries were incubated at 39 ℃ in the media contained 1.5 mg/ml collagenase for 60 min to disperse the intact preantral follicles from the ovarian stroma. Trial I was undertaken to investigate the effect of the different stage of development of preantral follicle on the growth and survival of its oocyte in vitro. The isolated preantral follicles at different stage of development [classified by oocyte diameter into <70 μ m (small) 70 ~ <80 μ m (middle) and 80 ~ 90 μ m (large)] were embedded in Ca-alginate gel and then were cultured in growth media [Waymouth MB 752/1 medium (95%) + FCS (5%) + FSH (2 μ g/ml) + E �� (1 μ g/ml)] for 16 days. The survival of oocytes assayed by FDA (3',6'- diacetyl fluorescin) staining technique and the growth of oocytes in diameter in preantral follicles were evaluated in each 4 days. During cultivation, the size in diameter increased from 260, 320 and 390 μ m (Day 0) to 520, 600 and 610 μ m (Day 16) for follicles, and from 58, 71 and 84 μ m (Day 0) to 86, 106 and 114 μ m (Day 16) for oocytes for the small, middle and large groups, respectively. At the end of cultivation, the forming rate of antrum cavity of follicle of three groups were 0%, 4% and 14%, respectively, and the survival rate of oocyte decreased significantly, from 39, 53 and 63% on Day 4 to 10, 28 and 25% on Day 16, respectively. The growth and the survival of oocytes during cultivation were affected by the initial oocyte size of preantral follicles. The miiotic competence ofoocytes grown in vitro [from cultured larger preantral follicles (80-90 μ m oocyte diameter) for 12 days] was assessed and compared with that of oocytes grown in vivo [from ovarian follicles (>1 mm in diameter) ] in maturation cultivation for 44-48 hr in trial II. After cultivation, the maturation rate (progressed to metaphase II) of the grown oocytes (>90 μ m in diameter) in vivo were significant higher than that oocytes grown in vitro [61% (169/274) vs. 2% (2/103), p<0.05]. All of the smallest oocytes (<90 μ m in diameter) grown both in vitro and in vivo, should remain at germinal vesicle stage after maturation cultivation. Results of the experiments revealed that the oocytes of preantral follicles at different stages of development affected significantly the survival of oocytes and the growth of follicles and oocytes in diameter in vitro during 16 days growth cultivation. The survival of preantral follicular oocyte decreased significantly during cultivation in vitro. After maturation cultivation, the meiotic competence of the grown oocytes both in vitro and in vivo were increased progressively with the final growth of oocyte in diameter. |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。