Traditionally, cell culture medium in iPSC-derived cell work is not the main focus of the research and often is considered as just "food for cells". We demonstrate that by manipulation of the media and optimized methodology, it is possible to use this solution to study the proteins that the cell secretes (the "secretome"). This is particularly useful in the study of iPSC-derived neurons, which require long culture time. We demonstrate that media can be used to model diseases with optimized incubation and sampling times. The ability not to sacrifice cells allows significant cost and research benefits. In this manuscript we describe an optimized method for the analysis of the cell media from iPSC-derived neuronal lines from control and Parkinson's disease patients. We have evaluated the use of standard and supplement B27-free cell media as well as five different sample preparation techniques for proteomic analysis of the cell secretome. Mass spectral analysis of culture media allowed for the identification of >500 proteins, in 500 μL of media, which is less volume than reported previously (20-40 mL). Using shorter incubation times and our optimized methodology, we describe the use of this technique to study and describe potential disease mechanisms in Parkinson's disease.

中文翻译：

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一种用于蛋白质组学分析的少量细胞培养基和分泌组的优化方法：帕金森氏病患者iPSC衍生的神经元细胞系中蛋白质表达改变的应用和验证。

传统上，iPSC衍生的细胞工作中的细胞培养基不是研究的重点，通常被视为“细胞的食物”。我们证明，通过操纵培养基和优化方法，可以使用这种解决方案来研究细胞分泌的蛋白质（“秘密组”）。这在需要较长培养时间的iPSC衍生神经元的研究中特别有用。我们证明，可以使用培养基来优化疾病的孵化和采样时间。不牺牲细胞的能力带来了可观的成本和研究收益。在此手稿中，我们描述了一种优化方法，用于分析来自对照和帕金森氏病患者的iPSC来源的神经元系的细胞培养基。我们已经评估了使用标准和无B27补充细胞培养基以及五种不同的样品制备技术对细胞分泌蛋白组进行蛋白质组学分析的方法。培养基的质谱分析可以鉴定500μL培养基中的> 500种蛋白质，其体积比以前报道的要少（20-40 mL）。使用较短的孵育时间和我们优化的方法，我们描述了使用该技术来研究和描述帕金森氏病的潜在疾病机制。