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題名 | 鰻魚肝細胞之懸浮條件探討 |
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作者 | 黃登福; 林瑞芬; 鄭森雄; | 書刊名 | 臺灣水產學會刊 |
卷期 | 21:3 1994.09[民83.09] |
頁次 | 頁273-280 |
分類號 | 387.108 |
關鍵詞 | 分離肝細胞; 鰻魚; 懸浮條件; Isolated liver cell; Eel; Suspending condition; |
語文 | 中文(Chinese) |
中文摘要 | 為建立魚類肝細胞作為毒理和代謝等研究之體外(in vitro)實驗素材,乃利用 灌流方式先清除鰻魚肝臟血液,再利用膠原蛋白消化肝臟結締組織而製得鰻魚肝細胞。分離 所得之鰻魚肝細胞,經探討不同懸浮條件如溫度、肝細胞濃度和懸浮液組成對肝細胞存活率 之影響,得知為90%以上活率之鰻魚肝細胞,在已試之條件中,其在懸浮液Krebs-HEPES buffer中之較佳懸浮溫度為37℃,時間為2小時;肝細胞在懸浮液之較佳濃度為2×107 cells/ml,懸浮液之較佳pH值為7.5。其次懸浮液中添加5.6mM葡萄糖和2.5mM鈣離子均對 鰻魚肝細胞之體外活存率具有提昇之效用。 |
英文摘要 | To establish fish hepatocytes as experimental materials for in vitro system of toxicological and metabolic studies, the livers of Japanese eel Auguilla japonica were used to isolate hepatocytes by using perfusion method. Based on the viability of hepatocytes by staining with trypan blue the optimal suspending conditions for eel hepatocytes were determined. It was found that the optimal suspending temperature, time, cell concentration and pH value were 37℃, 2 hr x 107 cells/ml and 7.6, respectively. It was also found that the viability of isolated eel hepatocytes was improved when the suspending buffer was added with either 5.6 mM glucose or 2.5 mM Ca++ |
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