HDL-Mediated Cellular Cholesterol Efflux Assay Method.
HDL-Mediated Cellular Cholesterol Efflux Assay Method.
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发表时间：
2015-11
影响因子：
0.8
通讯作者：
A. Hafiane;J. Genest
中科院分区：
文献类型：
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作者：
A. Hafiane;J. Genest
高密度脂蛋白（HDL）功能的生物标志物可能提供机制的见解和更好的心血管风险的歧视比HDL-胆固醇质量。这项工作的目的是描述一个简化的实验方案，可用于测定胆固醇流出从巨噬细胞培养的细胞，并被带到一个中等的吞吐量。细胞胆固醇流出试验旨在量化胆固醇从培养细胞流出到受体颗粒或血浆的速率。该测定是多步骤的、基于细胞的测定。如果不仔细控制，各种因素可能会影响测定的准确度和重现性。尝试解决影响该测定的因素，并提供相对快速和可扩展的标准化方法。我们证明，进一步离心的HDL组分是必要的，以避免载脂蛋白B污染时，使用聚乙二醇（PEG）的方法。我们还证明，当使用PEG与血浆或血清时，对胆固醇流出效率没有影响。该方法以前已在我们的实验室中应用于心血管研究，心血管疾病和药物治疗。
Biomarkers of high-density lipoprotein (HDL) function may provide mechanistic insights and better cardiovascular risk discrimination than HDL-cholesterol mass. The purpose of this work is to describe a simplified experimental protocol that can be used in the determination of cholesterol efflux from macrophages cultured cells and be brought to a medium throughput volume. The cellular cholesterol efflux assay is designed to quantify the rate of cholesterol efflux from cultured cells to an acceptor particle or to plasma. This assay is multi step, cell based assay. Various factors, if not carefully controlled may influence the accuracy and reproducibility of the assay. Attempts were made to address factors influencing this assay and to provide a standardized method that is relatively rapid and scalable. We demonstrate that further centrifugation of the HDL fraction is necessary to avoid apolipoprotein B contamination when using polyethylene glycol (PEG) method. We demonstrate also no effect on cholesterol efflux efficiency when using PEG with plasma or serum. This method has been previously applied in our laboratory in context of cardiovascular research, cardiovascular disease and pharmacologic therapies.