José A A F B V, Gama M A S, Urban A, Merighe G K, Meirelles F V, Etchegaray M A L, Lanna D P D
Departamento de Zootecnia, Universidade de São Paulo, Piracicaba, SP, Brasil.
Genet Mol Res. 2007 Mar 30;6(1):214-21.
Cultures of adipose tissue explants are a valuable tool for studying the intracellular mechanisms involving hormones and nutrients. However, testing how fatty acids affect cells requires a carrier molecule; bovine serum albumin (BSA) has been used for this purpose. However, contaminants can alter the cellular response. Our objectives were to: 1) test BSA as a fatty acid carrier and 2) evaluate polyvinyl alcohol (PVA) as a replacement for BSA. Adipose tissue explants from nine pigs were cultured in medium 199 for 4, 12, 24, and 48 h, with the following treatments: control, PVA (100 mM PVA added) and PVA + pGH (100 mM PVA plus 0.1 mg/mL porcine growth hormone). After each culture period, explants were collected and assayed for lipogenesis. After 48 h in culture, explants were assayed for lipolysis. A preliminary study with different commercial sources and high concentrations showed that BSA affected lipogenic rates. On the other hand, there were no effects of PVA on lipid synthesis, while pGH (positive control) reduced glucose incorporation into lipids (P < 0.01) when compared to both control and PVA (P < 0.05). There was no difference between control and PVA for lipolysis rates. However, pGH increased lipolysis when compared to control (P < 0.01) and PVA (P < 0.05). We demonstrated that BSA can alter lipogenesis, which precludes its use as a carrier molecule. On the other hand, addition of PVA had no effect on lipolysis or lipogenesis. We suggest the use of PVA instead of BSA for adding bioactive fatty acids to cultures of adipose tissue.
脂肪组织外植体培养是研究涉及激素和营养物质的细胞内机制的一种有价值的工具。然而,测试脂肪酸如何影响细胞需要一种载体分子;牛血清白蛋白(BSA)一直被用于此目的。然而,污染物会改变细胞反应。我们的目标是:1)测试BSA作为脂肪酸载体,2)评估聚乙烯醇(PVA)作为BSA的替代品。将来自9头猪的脂肪组织外植体在199培养基中培养4、12、24和48小时,采用以下处理:对照、PVA(添加100 mM PVA)和PVA + pGH(100 mM PVA加0.1 mg/mL猪生长激素)。在每个培养期后,收集外植体并测定脂肪生成。培养48小时后,对外植体进行脂肪分解测定。一项使用不同商业来源和高浓度的初步研究表明,BSA会影响脂肪生成率。另一方面,PVA对脂质合成没有影响,而与对照和PVA相比,pGH(阳性对照)降低了葡萄糖掺入脂质的量(P < 0.01)(P < 0.05)。对照和PVA的脂肪分解率没有差异。然而,与对照(P < 0.01)和PVA(P < 0.05)相比,pGH增加了脂肪分解。我们证明BSA会改变脂肪生成,这使其不能用作载体分子。另一方面,添加PVA对脂肪分解或脂肪生成没有影响。我们建议在向脂肪组织培养物中添加生物活性脂肪酸时使用PVA而不是BSA。