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題 名 | 凍解及新鮮豬黃體細胞對激黃體物質於體外分泌孕酮之效應=Responses of Progesterone Secretion in Vitro by Defrozen and Fresh Porcine Luteal Cells to Luteotropic Compounds |
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作 者 | 曾啟明; 林仁壽; | 書刊名 | 中國畜牧學會會誌 |
卷 期 | 27:2 1998.06[民87.06] |
頁 次 | 頁229-242 |
分類號 | 437.21 |
關鍵詞 | 豬; 凍解黃體細胞; 激黃體物質; 孕酮; 細胞培養; Pig; Defrozen luteal cell; Luteotropic compound; Progesterone; Cell culture; |
語 文 | 中文(Chinese) |
中文摘要 | 採自桃園母豬於動情週期第12天之黃體,應用酵素解離並將大部份黃體細胞(LC) 凍存於液態氮桶中,供探討凍解(冷凍-解凍)及新鮮LC對不同劑量之排卵素(oLH)、環腺甘 單磷酸(cAMP)、或人類絨毛膜激性腺素(hCG)之孕酮(progesterone)分泌效應,俾建立一套豬 凍解LC的體外培養係統。由結果顯示,每gm黃體組織經解離後可獲得3.2�e10 個新鮮LC, 存活率為84.3%;而凍解LC經培養24hr之存活率則降為20~25%。由凍解LC之孕酮分泌 時程反應顯示,當凍解LC數目為10 /mL時,縱使不添加22R-羥膽固醇作為受質,亦可 獲良好激黃體效應。再者,孕酮生產速率於培養4~6hr期間大幅揚升,及至12~24hr期間則 高原態。凍解LC對oLH、cAMP和hCG的刺激孕酮產量雖為同一來源之新鮮LC者的 1/15~1/20,惟均呈劑量反應,其呈激黃體效應(P<0.05)的最低劑量分別為100ng/mL、10�嵱 或0.01IU/mL。此一結果酷似新鮮LC者;尤其凍解與新鮮LC仍保有大部份原新鮮LC對 激黃體物質刺激而分泌孕酮之特性孕酮之特性。綜前所述,本研究所建立的豬LC於凍解後, 採用10 /mL個細胞經6hr或24hr培養,對激黃體效應均可呈理想結果。 |
英文摘要 | This study was designed to establish an in vitro culture system for frozen-thawed porcine luteal cells. Corpora lutea were collected from Toayuan sows on day 12 of the estrous cycle and dissociated enzymically to obtain the dispersed luteal cells (LC). Most portion of the LC were cryopreserved in a liquid-nitrogen tank. Ovine luteinining hormone (oLH), cyclic adenosine monophosphate (cAMP) or human chorionic gonadotropin (hCG) was used to determine the dose- dependent responsiveness of progesterone (P ) secretion by the fresh (FLC) and defrozen (DLC) luteal cell. The results showed that the number and viability of the harvested FLC were 3.2�e10 /gm tissue and 84.3%, respectively, while the viability of DLC decreased to 20~25% by 24 hr after incubation. The time-course responsiveness showed that when the number of DLC incubated was 10 /mL, a good result for luteotropic effect could be obtained even though a soluble hydroxycholesterol analogue (22R-OHC) was not supplemented as a substrate. Moreover, P accumulation of DLC to luteotropic compounds increased promptly during the 4~6 hr incubation and went up to a plateau during the 12~24 hr incubation. Additionally, each of the luteotropic compounds stimulated P production from DLC in a dose-dependent manner although the amount of P production from DLC was 1/15~1/20 of that from FLC during the 24 hr incubation. The lowest effective doses for luteotropic response to oLH, cAMP and hCG were 100 ng/mL, 10�嵱 and 0.01IU/mL, respectively. These results were very similar to FLC ones. Especially, the profiles in dose response of P production from both DLC and FLC to cAMP stimulation were quite the same, suggesting that defrozen LC held most part of the luteotropic characteristics of original fresh one. These findings demonstrated that a culture system in vitro for frozen-thawed luteal cells in pig was established. A better responsiveness of P secretion to luteotropic compounds may be obtained at 6 or 24 hr after incubation with 10 /mL of defrozen luteal cells. |
本系統中英文摘要資訊取自各篇刊載內容。