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題名 | 豬卵巢組織浸於膠原蛋白酶溶液之時間和包埋處理對離體培養之腔前濾泡內卵母細胞之生長與存活率之影響=Effects of the Time for Incubation of Minced Porcine Ovary in a Solution Containing Collagenase, and the Embedding of Preantral Follicles with Various Coating Matrix, on the Growth and Survival of Oocytes Cultured in Vitro |
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作者 | 顏大慶; 鄭三寶; 劉炳燦; 顏淑卿; 邱啟明; | 書刊名 | 中國畜牧學會會誌 |
卷期 | 26:2 1997.06[民86.06] |
頁次 | 頁197-210 |
分類號 | 437.21 |
關鍵詞 | 腔前濾泡; 豬卵母細胞; 膠原蛋白酶; 包埋材質; 體外存活率; Preantral follicle; Porcine oocyte; Collagenase; Coating matrix; Survival in vitro; |
語文 | 中文(Chinese) |
中文摘要 | 本試驗旨在探討豬卵巢組織浸於膠原蛋白�t (Collagenase)溶液之時間和培養前 之包埋處理, 對腔前濾泡內之卵母細胞於離體短時程 (4 天 ) 之發育及其存活率之影響。 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。結果顯示,於所收集之僅由卵丘細胞包被或已裸露之卵母細胞,以及腔狀或腔 前濾泡內卵母細胞之存活率, 不因已細切之卵巢組織被浸於含 1.5 mg/ml 膠原蛋白�t溶液 中經歷時間之不同 (30、45、60 和 75 min) 而有所影響 (P>0.05)。 腔前濾泡經海藻酸鈣 膠層包埋並置于成長培養液 [Waymouth MB752/1 (95 % ) + FCS (5 % ) + FSH (2 μ g/ml) + E �� (1 μ g/ml)] 中培養 4 天, 則培養前後卵母細胞之直徑, 於 60 和 75 min 兩組間之差異不顯著; 惟 75 min 組之卵母細胞之存活率顯著低於 60 min 組者 (30 % vs. 60 %, P<0.05)。 此結果啟示, 已細切之豬卵巢組織被浸於膠原蛋白�t溶液 (1.5 mg/ml) 中經歷超過 60 min 之處理, 雖有助於隨後應用機械剝離以收集腔前濾泡之數目, 惟將不利於腔前濾泡內卵母細胞之生物活性。在評估腔前濾泡於培養前使用不同材質包埋處 理之實效顯示,於離體生長培養前後之卵母細胞之直徑不因包埋組別之不同而受影響;惟於 培養後,應用海藻酸鈣膠層所包埋之卵母細胞,其存活率顯著高於應用膠原蛋白膠層包埋者 或未予包埋者 (52 % vs. 30 %或 18 %, P<0.05)。 |
英文摘要 | The purpose of this study was to evaluate the effects of the time for incubation of minced porcine ovary in a collagenase solution, and the embedding of isolated preantral follicles with various coating matrix, on the growth and survival of oocytes during short-time cultivation (4 days) in vitro. Trial I was conducted to investigate the effect of time (30, 45, 60 and 75 min, respectively) of incubation in the media containing 1.5 mg/ml collagenase of ovary tissues on the survival of the oocytes collected from the cumulus-oocytes complex, the nude oocytes and the preantral and antral follicles. The survival rates of collected oocytes [assayed by FDA (3',6'- diacetyl fluorescin) staining technique ] were not different among incubation time groups (P>0.05). Therefore, the isolated follicles collected from the 60 min- and 75 min- incubation time in collagenase solution of ovary tissues were embedded in Ca-alginate gel and then cultured in growth media [Waymouth MB752/1 (95%) + FCS (5%) + FSH (2 μ g/ml) +E �� (1 μ g/ml)] for 4 days. After cultivation, there were differences in survival rate of oocytes in preantral follicles between 75 min-incubation group and 60 min-incubation group (30% vs. 60%, P<0.05), but was not in the diameter of oocytes. In trail II, the preantral follicles were embedded in different coating matrix before cultivation. The growth of oocytes in diameter before and after cultivation were not different among groups (Ca-alginate gel, collagen gel and uncoating control), but the survival rate of oocytes embedded in Ca-alginate gel after cultivation was significantly higher than the oocytes embedded in collagen gel or in uncoating control (52% vs. 30% or 18%, P<0.05). It indicated that the coating matrix with Ca-alginate will be useful to maintain the survival of oocytes in preantral follicle during cultivation |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。