Preparation of Lipid-Stripped Serum for the Study of Lipid Metabolism in Cell Culture
Preparation of Lipid-Stripped Serum for the Study of Lipid Metabolism in Cell Culture
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DOI：
10.21769/bioprotoc.2876
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发表时间：
2018-06-05
期刊：
影响因子：
0.8
通讯作者：
Vander Heiden, Matthew G.
中科院分区：
文献类型：
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作者：
Hosios, Aaron M.;Li, Zhaoqi;Vander Heiden, Matthew G.
研究培养细胞中的脂质代谢很复杂，因为细胞通常是在动物血清存在的情况下培养的，动物血清中含有广泛、可变且不确定的脂质种类。脂质代谢可以影响细胞生理学、信号传导和增殖，并且在没有外源脂质的情况下培养细胞的能力可以揭示脂质生物合成途径的重要性并促进具有特定脂质种类的培养基的生成。我们采用了一种方案，从血清中去除脂质，而不消除其支持培养细胞增殖的能力。该方法需要二异丙醚和丁醇，可用于在四天内生成小批量的去脂血清。所得血清支持培养物中许多细胞系的增殖，并可用于比较细胞在脂质充足和耗尽条件下的代谢。
Studying lipid metabolism in cultured cells is complicated by the fact that cells are typically cultured in the presence of animal serum, which contains a wide, variable, and undefined variety of lipid species. Lipid metabolism can impact cell physiology, signaling, and proliferation, and the ability to culture cells in the absence of exogenous lipids can reveal the importance of lipid biosynthesis pathways and facilitate the generation of media with defined lipid species. We have adapted a protocol to remove lipids from serum without eliminating its ability to support the proliferation of cells in culture. This method requires di-isopropyl ether and butanol and can be used to generate small batches of lipid-stripped serum in four days. The resulting serum supports proliferation of many cell lines in culture and can be used to compare the metabolism of cells in lipid replete and depleted conditions.