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頁籤選單縮合
題 名 | 豬胚的體外生產=In Vitro Production of Porcine Embryos |
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作 者 | 吳明哲; 李秀美; | 書刊名 | 畜產研究 |
卷 期 | 31:1 1998.03[民87.03] |
頁 次 | 頁53-67 |
分類號 | 437.21 |
關鍵詞 | 豬; 卵; 胚; 體外成熟; 體外受精; 體外培養; Pig; Oocyte; Embryo; IVM; IVF; IVC; |
語 文 | 中文(Chinese) |
中文摘要 | 豬卵自屠宰肉豬卵巢之成長濾泡收集,經體外成熟與受精後,再予以體外培養至 八細胞胚期之過程,是豬胚體外生產的操作工作。試驗一:第一日下午三點起對豬死後一小時 左右的卵巢,進行濾泡卵的回收,有卵丘細胞包被的卵才予以體外成熟培養,下午四點置放 於50μl的體外成熟培養小滴內,在有5%二氧化碳供應狀態的39℃恆溫培養箱培養41小時。 第三日早上九點前洗滌體外受精所需的精子並調整為2x10μ精子/ml濃度備用。把每10個卵 置放於含有咖啡因的體外受精用培養小滴中,加50μl等量的稀釋精液後放入培養箱培養6 小時。到下午三點取出,洗去卵丘細胞層外多餘的精子,再置放於體外培養小滴中繼續培養 42小時或66小時,亦即第五日或第六日的上午九點檢視細胞分裂狀況。有89.8%(3640/4050) 的收集卵是有卵丘細胞包被者。整個體外生產過程中計有1,898個卵受到污染,污染發生率 為52%(1898/3640)。在加入精子後48小時的652個卵分別有1.38%和3.53%的卵發育為雙細 胞胚和四細胞胚;而於72小時後的1,090個卵分別有1.01、2.20和4.04%的卵發育成雙細胞 胚、四細胞胚和八細胞胚。試驗二:體外生產流程如試驗一,但改以100個卵置入3 ml培養 液中為一培養單位,胚期評估時間為加入精子後的50小時。計有2180個卵被培養,污染發生 率為28%(600/2180)。卵於加入精子後50小時,體外生產胚的產製率有15.38%(243/1580)。 發育的243個胚分別有28.81、32.92和38.27%為雙細胞胚、四細胞胚和八細胞胚。 |
英文摘要 | In vitro production (IVP) of porcine embryos consists of collection of follicular oocytes, in vitro maturation of oocytes, in vitro fertilization, and in vitro culture of ertilized oocytes up to the 8-cell embryo stage. In Exp. 1, experiments were begun at 3 p.m. of the first day of IVP by the collection of follicular oocytes from ovaries of gilts one hour postmortem, and subsequently the cultivation of them by 4 p.m. in a 39℃ cell incubator with supplement of 5% CO2 in air for 41 hours. By 9 a.m. of the third day of IVP, sperm suspension had been washed and also adjusted to the concentration of 2 X 10□ sperm/ml. Groups of 10 oocytes were prepared into a droplet of fertilization medium supplemented with caffeine, and then added to them an equal volume of 50μl of semen suspension. After an in vitro fertilization period of six hours, loosely binding sperm were washed away from the cumulus cells around the oocyte. Oocytes were then transferred into a droplet of culture medium at 3 p.m. of the third day of IVP. After 42 or 66 hours of in vitro culture, the development of oocytes were examined either at 9 a.m. of the fifth or sixth day of IVP. A total of 4050 follicular oocytes was recovered, but only 89.8% (3640/4050) of them were used for in vitro maturation by the characteristics of encompassed cumulus cells around the oocytes. During the processes of IVP, 1898 oocytes were contaminated which represented 52% (1989/3640) of contamination rate. By 48 hours after insemination, 1.38 and 3.53% of 652 oocytes developed into 2-cell embryo and 4-cell embryo, respectively. Of the 1090 oocytes, 1.01, 2.20 and 4.04% were respectively developed into 2-cell, 4-cell or 8-cell embryos by 72 hours after insemination. In Exp. 2, protocols were used as described in Exp. 1 except that a culture unit consisted of 100 oocytes and examination on embryo cleavage was taken at 50 hours after insemination. A total of 2180 oocytes was used to culture, and 28% (600/2180) of contamination rate was observed. Production rate of porcine embryos produced in vitro was 15.38% (243/1580) at 50 hours after insemination. Of the 243 embryos developed, 28.81, 32.92 and 38.27% of embryos were 2-cell, 4-cell and 8- cell embryos, respectively. |
本系統中英文摘要資訊取自各篇刊載內容。