The effect of culture on morphology, lipid and fatty acid composition, and polyunsaturated fatty acid metabolism of rainbow trout (Oncorhynchus mykiss) skin cells
The effect of culture on morphology, lipid and fatty acid composition, and polyunsaturated fatty acid metabolism of rainbow trout (Oncorhynchus mykiss) skin cells
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DOI：
10.1023/a:1007709508786
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发表时间：
1997-11-01
影响因子：
2.9
通讯作者：
Sargent, JR
中科院分区：
文献类型：
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作者：
Ghioni, C;Tocher, DR;Sargent, JR
虹鳟鱼（Oncorhynchus mykiss）皮肤细胞培养物通过胰蛋白酶消化组织获得，并在Leibovitz L-15培养基中生长。脂质类组合物，总脂质和单个磷脂类的脂肪酸谱在不同时间的文化。通过在7天和14天后用[1-C-14]18：2n-6和[1-C-14-]18：3 n-3孵育原代培养物来研究多不饱和脂肪酸（PUFA）的代谢。上皮样原代培养物和成纤维细胞样次级继代培养物之间的形态变化伴随着脂质组成的改变。极性脂质在培养14天后占主导地位。磷脂酰胆碱（PC），最丰富的磷脂，磷脂酰肌醇和胆固醇的相对比例显着增加，而鞘磷脂减少。饱和脂肪酸，18：1 n-9，n-6和n-9 PUFA更丰富的总脂质在培养物中，在14天和4个月比最初分离的细胞，其中含有较高的百分比的长链单烯和n-3 PUFA。在所有主要磷脂类中观察到脂肪酸组成随培养时间的变化。培养的虹鳟鱼皮肤细胞在18：2n-6和18：3 n-3两种情况下都去饱和并伸长，其中20：4 n-6和20：5 n-3分别是最丰富的产物。PC表现出最高的放射性掺入，尤其是在与18：3 n-3孵育后。脂质代谢一般随着原代培养物的年龄而增加，与7天培养物相比，14天培养物中通过去饱和/伸长掺入和代谢的CLX PUFA的量显著增加。计算的n-6和n-3脂肪酸的产物/前体比表明，虽然Delta 6去饱和酶活性随细胞年龄显著增加，但Delta 5去饱和酶活性更受脂肪酸系列的影响，18：3 n-3比18：2n-6更容易转化为20：4 n-6。20：5 n-3进一步去饱和生成六烯的反应很低。总体而言，数据表明，鳟鱼皮肤细胞培养物比哺乳动物表皮/角质形成细胞培养物更类似于哺乳动物皮肤成纤维细胞。
Rainbow trout (Oncorhynchus mykiss) skin cell cultures were obtained by trypsinization of the tissue and grown in Leibovitz L-15 medium. Lipid class compositions, and fatty acid profiles of total lipids and individual phospholipid classes were determined at different times of culture. The metabolism of polyunsaturated fatty acids (PUFA) was investigated by incubating primary cultures after 7 and 14 days with [1-C-14]18:2n-6 and [1-C-14-]18:3n-3. The change in morphology between epithelial-like primary cultures and fibroblastic-like secondary subcultures was accompanied by alterations in the lipid composition. Polar lipids became predominant by 14 days in culture. The relative proportions of phosphatidylcholine (PC), the most abundant phospholipid, phosphatidylinositol and cholesterol increased significantly, while sphingomyelin decreased. Saturated fatty acids, 18:1n-9, n-6 and n-9PUFA were more abundant in total lipid in cultures at 14 days and 4 months than in cells initially isolated which contained higher percentages of longer chain monoenes and n-3PUFA. The changes in fatty acid composition with time in culture were observed in all the major phospholipid classes. Rainbow trout skin cells in culture desaturated and elongated both 18:2n-6 and 18:3n-3, with 20:4n-6 and 20:5n-3 being the most abundant products, respectively. PC presented the highest incorporation of radioactivity, especially following incubation with 18:3n-3. Lipid metabolism in general increased with the age of primary cultures, with both the amount of CLX PUFA incorporated and metabolized by desaturation/elongation significantly increased in 14 day cultures compared to 7 day cultures. Product/precursor ratios calculated for both n-6 and n-3 fatty acids showed that, while Delta 6 desaturase activity was increased significantly with cell age, Delta 5 desaturase activity was more affected by the fatty acid series, with 18:3n-3 being more readily transformed to 20:5n-3 than 18:2n-6 to 20:4n-6. Further desaturation of 20:5n-3 to hexaenes was low. Overall, the data suggested that the trout skin cell cultures were more similar to mammalian skin fibroblasts than mammalian epidermal/keratinocyte cultures.