Effects of albumin bound fatty acids on the growth of the human T lymphoblastic cell line Jurkat
Effects of albumin bound fatty acids on the growth of the human T lymphoblastic cell line Jurkat
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白蛋白结合脂肪酸对人 T 淋巴细胞系 Jurkat 生长的影响
DOI：
10.1007/s11626-011-9454-z
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发表时间：
2011
影响因子：
2.1
通讯作者：
Takahashi S
中科院分区：
文献类型：
--
作者：
Okazaki T;Iwasaki T;Fukuoka A;Suzuki M;Katagiri H;Okano T;Takahashi S
为了在RPMI 1640培养基中培养T淋巴母细胞系JURKAT，加入人白蛋白而不是胎牛血清（FBS）至终浓度为1%，以促进与FBS相当的生长。此外，在培养物中加入几种动物白蛋白，导致生长速率明显不同（图1）。在RPMI 1640培养基中，包括氨基酸和碳水化合物作为营养物，但不包括脂质。大多数脂肪酸与血清白蛋白和甲胎蛋白结合（Reed 1986; Copado et al. 1999），白蛋白可能是本研究中培养细胞的唯一脂质来源。另一方面，已知脂肪酸对细胞生长具有多种功能;即，细胞毒性（Lima et al. 2002; Martins de Lima et al. 2006）、诱导细胞凋亡（Welters et al. 2004; Artwohl et al. 2008）和调节信号传导（Pawar and Jump 2003）。在此，我们使用测量试剂盒（和子，Tokyo，Japan）测量动物白蛋白（10%溶液）中非酯化脂肪酸（NEFA）的浓度，并使用气相色谱法分析白蛋白结合的NEFA。结合NEFA的量随白蛋白而变化，兔白蛋白最大，其次是羊、大鼠、人和牛白蛋白。FBS中的NEFA显示介于大鼠和人白蛋白之间的值（图2A）。使用气相色谱法，分析与动物白蛋白结合的脂肪酸由棕榈酸（C16：0）、硬脂酸（C18：0）和油酸（C18：1）或亚油酸（C18：2）组成，类似于用于Jurkat细胞培养的FBS中的NEFA（图2 B）。然而，只有兔白蛋白附着大量辛酸（C8：0），这可能导致纯化过程中的污染，绵羊白蛋白附着的硬脂酸（C18：0）量是大鼠或人白蛋白的两倍多（图2 C）。在补充有来自各种来源的白蛋白的培养基中，
To culture a T lymphoblastic cell line, JURKAT, in RPMI 1640 medium, human albumin, instead of fetal bovine serum (FBS), was added to a final concentration of 1% to facilitate growth comparable with FBS. In addition, several kinds of animal albumin were added to the culture, resulting in markedly different growth rates (Fig. 1). InRPMI 1640 medium, amino acids and carbohydrates are included as nutrients, but no lipids. Most fatty acids are binding to serum albumin and alpha fetoprotein (Reed 1986; Copado et al. 1999), and albumin may be the only source of lipids for cultured cells in this study. On the other hand, it is known that fatty acids have various functions to cell growth; ie, cytotoxicity (Lima et al. 2002; Martins de Lima et al. 2006), induction of apoptosis (Welters et al. 2004; Artwohl et al. 2008), and regulation of signaling (Pawar and Jump 2003). Here, we measured the concentrations of non-esterified fatty acids (NEFAs) in animal albumin (10% solution) using a measuring kit (Wako, Tokyo, Japan), and analyzed albumin-bound NEFAs employing gas chromatography. The amounts of bound NEFAs varied with albumins, being largest for rabbit albumin, followed by sheep, rat, human, and bovine albumins in this order. NEFAs in FBS showed values intermediate between rat and human albumins (Fig. 2 A). Using gas chromatography, fatty acids bound to animal albumins were analyzed to be consisting of palmitic (C16: 0), stearic (C18: 0), and oleic (C18: 1) or linoleic acids (C18: 2) similar to NEFAs in FBS used for Jurkat cell culture (Fig. 2 B). However, only rabbit albumin attached a large amount of caprylic acid (C8: 0), which presumably caused contamination during the purification process, and sheep albumin attached more than twice as large amount of stearic acid (C18: 0) as to rat or human albumin (Fig. 2 C). In media supplemented with albumins derived from various