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題名 | 成熟培養液之濾泡液含量和受精處理對誘發豬卵母細胞產生孤雌生殖致活之影響=The Effect of Follicular Fluid Supplementation in Maturation Medium and Fertilization Treatment on the Parthenogenetic Activation of Porcine Oocytes |
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作者 | 黃尉東; 朱志成; 鄭三寶; 吳信志; 唐品琦; 劉炳燦; | 書刊名 | 中國畜牧學會會誌 |
卷期 | 28:3 1999.09[民88.09] |
頁次 | 頁359-372 |
分類號 | 437.21 |
關鍵詞 | 孤雌生殖; 豬; 卵子; 濾泡液; Parthenogenesis; Pigs; Oocytes; Follicular fluid; |
語文 | 中文(Chinese) |
中文摘要 | 本研究旨在評估豬卵母細胞於體外成熟與體外受精培養過程中,發生孤雌生殖致 活之百分率。自卵巢表面之腔狀濾泡(直徑≧3 mm)所取得之卵丘-卵母細胞複合體,逢機置於 含有不同比例濾泡液之基礎成熟培養液[TCM-199+FSH(2.5μg/ml)+LH(5 IU/ml)]中培養 44h,經與體外獲能之精子於m199-NBCS受精培養液中混合培養8~10h後,始移入含20% porcine serum(PS,day 0)之mBMOC-2培養液中繼續後續培養迄96h。 結果顯示:卵母細胞經體外成熟培養後,進行未加入活精子之受精培養(8~10h),後轉入 mBMOC-2+20% PS液中進行後續培養,其孤雌生殖致活率自後續培養8~10h之0~3.8%增加至 後續培養96h之18.5~29.2%。若在受精培養期間,將卵母細胞仍留置于成熟培養液時,其孤 雌生殖致活率與移入m199-NBCS受精培養液中進行受精培養者相近。然而,二者之孤雌生殖 致活率均隨成熟培養液中濾泡液含量之增加而增加;其中,100%濾泡液組者顯著高於0%或20% 濾泡液組者(29.3% vs. 18.5或18.7%, P<0.05)。 當卵母細胞進行體外成熟後與體外獲能之精子進行受精處理時,于20,40和100%等三種 濾泡液含量組之受精卵分裂發育率,分別為24.8,34.3和30.1%,而未與精子進行受精處理之 孤雌生殖致活率,則分別為16.2,16.4和13.6%。因此估計,卵母細胞之孤雌生殖致活率(未 受精處理組)約為受精卵分裂發育率(受精處理組)之50~60%。惟受精處理組之受精卵分裂發 育率,或未經受精處理組之孤雌生殖致活率,於三種濾泡液含量組別間之差異均不顯著 (P>0.05)。 上述結果顯示,卵母細胞被置于含濾泡液之TCM-199液中歷經成熟培養,或/和受精培養 後,引發孤雌生殖致活之機率,隨受精培養後之後續培養時間之增長而增加。於本試驗中, 更換受精培養液並未對其發生率有促進或抑制作用;然而,當成熟培養液中之濾泡液含量增加 時,則對孤雌生殖致活之發生率有促進趨勢。此外,就未與精子進行受精處理所得之資訊, 評估卵母細胞歷經成熟培養並與獲能精子進行受精處理後之分裂發育胚中,半數可能係源自 卵母細胞自身發生孤雌生殖致活之結果。 |
英文摘要 | The aim of this experiment was to investigate the activation of parthenogenesis during the process of in vitro maturation (IVM) and fertilization (IVF) of porcine oocytes, and to evaluate the percentage of parthenogenetic embryos after IVM and IVF treatments. Porcine cumulus-oocyte complexes (COCs) were collected from the ovarian follicles (≧3 mm) of gilt. COCs were cultured in a basal maturation medium (TCM-199+FSH 2.5 μg/ml+LH 5 IU/ml) supplemented with various levels (0, 20, 40 and 100%) of porcine follicular fluid (pFF) for 44 h and for another 8-10 h in modified M199-New born calf serum (mM199-NBCS) with or without capacitated spermatozoa. The oocytes were then transferred to modified BMOC-2 medium (mBMOC-2) plus 20% porcine serum (PS, day 0) and incubated for 96 h. All the incubations were conducted in an environment containing 5% CO2 in humidified air at 39℃. Results indicated that when IVM oocytes were exposed under no sperm condition, and were cultured in maturation medium or transferred to mM199-NBCS for 8-10 h, their parthenogenesis rates were 0-3.8%. When subsequently transferred to mBMOC-2+20% PS for another 96 h, the parthenogenesis rates increased to 18.5-29.2%. It showed that the percentages of parthenogenesis embryos tend to be elevated when the culture period was prolonged. It was similar to those cultured in the original maturation medium or those moved to m199-NBCS for 8-10 h and for another 96 h. The percentage of parthenogenesis in 100% pFF-added medium was significantly higher than those in 0 and 20% pFF-added media (29.3% vs. 18.6 and 18.8%, P<0.05). This result indicated that the parthenogenesis seemed to be elevated as pFF levels increased. The cleavage rates of oocytes fertilized in pFF-added media (20, 40 and 100%) were 24.8, 34.3 and 30.1%, respectively, which were higher than those without fertilization treatment (16.2, 16.4 and 13.6%). The cleavage rate of unfertilized oocytes was 50-60% of those fertilized. The embryo cleavage rates of fertilized oocytes and parthenogenesis of those unfertilized were similar among these three pFF-added media (P>0.05). It is concluded that the possibility of parthenogenesis, in this study, was low (0-3.8%) when oocytes cultured in pFF-added TCM-199 medium and underwent subsequent fertilization treatment, but it increased when culture period was extended and possibly when pFF levels were increased during maturation. Replacement of the fertilization medium had no significant effect on the parthenogenesis of porcine oocytes. Half of the cleaved oocytes matured and fertilized in vitro might have originated from parthenogenesis. |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。