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題名 | 激黃體物質對山羊黃體細胞體外分泌孕酮之影響=Effects of Luteotropic Substances on Progesterone Secretion by Caprine Luteal Cells in Vitro |
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作者 | 吳兩新; 楊鎮榮; 黃政齊; 林仁壽; Wu, Leang-shin; Yang, Jenn-rong; Huang, Jan-chi; Lin, Jen-hsou; |
期刊 | 中國畜牧學會會誌 |
出版日期 | 19970900 |
卷期 | 26:3 1997.09[民86.09] |
頁次 | 頁313-326 |
分類號 | 437.58 |
語文 | chi |
關鍵詞 | 山羊; 黃體細胞; 孕酮; 細胞培養; Goat; Luteal cell; Progesterone; Cell culture; |
中文摘要 | 於臺灣山羊動情週期第9~12 天,以腹正中線剖腹術摘除卵巢剝取黃體,利用酵 素解離法 (含 collagenase 與 DNase I) 取得單離之黃體細胞。 解離後之黃體細胞進行細 胞存活率、直徑之測定及激黃體物質對其分泌孕酮 (progesterone) 能力之影響。結果顯示 , 每克黃體組織可得到 29.9 ± 9.3( × 10 �� ) 個細胞,解離後細胞存活率為 89.3 ± 7.9%;而經冷凍解凍後之存活率則為 86.1 ± 6.8%。 自組織切片測量大、小黃體細胞之直 徑平均分別為: 大細胞長徑為 28.7 ± 5.4um, 短徑為 21.6 ± 3.5um; 小細胞長徑為 17.7 ± 4.0um,短徑為 11.9 ± 2.5 um;另就單離之黃體細胞而言:大、小細胞直徑則分 別為 24.4 ± 3.4 和 16.7 ± 2.5um。 新鮮的山羊黃體細胞以激黃體物質刺激, 結果以 oLH 1g/ml 與 hCG 1 ng/ml 刺激 4 小時後, 其孕酮之分泌量即有顯著的增加 (P<0.05), 且孕酮之累積量至 12 小時為最高。 經冷凍解凍後之黃體細胞, 須在培養液中添加 22R-hydroxycholesterol 1 ug/ml 做為合孕酮之受質, 始能在激黃體物質的刺激下呈現劑 量反應, 而且以 oHL 100 ng/ml、hCG 1 ng/ml 之反應最好 (P<0.05), 並且在刺激後 24 小時孕酮的分泌量始有顯著之增加,孕酮累積量至 48 小時為最高。 |
英文摘要 | Ovaries were taken from Taiwan native goats on days 9~12 of estrous cysle by laparotomy. Corpus luteum (CL) was dissociated with enzyme digestion solution containing collagenase and DNase I, and then luteal cells were obtained. The dispersed luteal cells were used for determination of cell viability and diameter, as well as stimulation test of luteotropic substances in vitro. The results showed that the number of luteal cells harvested was 29.9 ± 9.3( × 10 �� )cells/g CL. The viability of fresh and frozen-thawed cells were 89.3 ± 7.9% and 86.1 ± 6.8%,respectively. The long and short diameter of the large luteal cells were 28.7 ± 5.1 and 21.6 ± 3.5 um on CL slice and small cells were 17.7 ± 4.0 and 11.9 ± 2.5 um, respectively. The average cell diameters of large and small single cells were 24.4 ± 3.4 and 16.7 ± 2.5um, respectively. In fresh luteal cells, a significant responsiveness of progesterone secretion was observed at oLH 1 ng/ml and hCG 1 ng/ml after 4 hours incubation (P<0.05), respectively, and the maximal accumulation of progesterone was not obtained until 12 hours. In frozen-thawed luteal cells, we found that the 22R-hydroxy cholesterol (22R-OHC) was an essential supplement for progesterone secretion. The optimal dose of 22R-OHC was 1 u g per ml. In exogenous luteotropic substances stimulation trials, a significant responsiveness of progesterone secretion was observed at oLH 100 ng/ml and hCG 1 ng/ml after 24 hours incubation (P<0.05), respectively, and the maximal accmulation of progesterone was not obtained until 48 hours. |
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