对复制衰老成纤维细胞模型中的脂质和蛋白质进行单细胞 MALDI-MSI 分析。

IF 3.1 2区 化学 Q2 BIOCHEMICAL RESEARCH METHODS
Emily R Sekera, Lorena Rosas, Joseph H Holbrook, Quetzalli D Angeles-Lopez, Timur Khaliullin, Mauricio Rojas, Ana L Mora, Amanda B Hummon
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引用次数: 0

摘要

在这项研究中,我们评估了人肺成纤维细胞(hLFs)中的脂质和特定蛋白质,以探究因衰老和衰老而发生的变化。为了研究单个细胞群,我们首先分析了使用聚赖氨酸和离心力沉积法粘附在载玻片上的细胞,以确定两种制备方法之间是否存在特定的变化。然后利用聚二赖氨酸方法检测细胞群的脂质体,并进一步评估 MALDI-免疫组化(IHC)平台在单细胞级分析中的潜在应用。在成纤维细胞群中观察到了两种衰老蛋白标记物--波形蛋白和 p21,并对其进行了量化。成纤维细胞的脂质组分析发现,12种脂质因复制衰老而发生了显著变化,包括脂肪酸(如硬脂酸)和神经酰胺磷脂醇胺物种(CerPE)。与之前的报告类似,在其他脂质种类中,也检测到了推定脂肪酸组成成分神经酰胺的改变。总之,我们的研究结果揭示了检测与衰老有关的脂质的能力,并显示了正常和衰老成纤维细胞群之间蛋白质表达的改变,包括年轻细胞和衰老细胞之间的差异。本报告是首次在单细胞水平上利用 MALDI-IHC 系统分析培养细胞中的蛋白质表达和脂质概况,尤其关注与衰老和衰老相关的变化。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Single Cell MALDI-MSI Analysis of Lipids and Proteins within a Replicative Senescence Fibroblast Model.

In this study, we evaluate lipids and select proteins in human lung fibroblasts (hLFs) to interrogate changes occurring due to aging and senescence. To study single cell populations, a comparison of cells adhered onto slides using poly-d-lysine versus centrifugal force deposition was first analyzed to determine whether specific alterations were observed between preparations. The poly-d-lysine approach was then utilized to interrogate the lipidome of the cell populations and further evaluate potential applications of the MALDI-immunohistochemistry (IHC) platform for single-cell-level analyses. Two protein markers of senescence, vimentin and p21, were both observed within the fibroblast populations and quantified. Lipidomic analysis of the fibroblasts found 12 lipids significantly altered because of replicative senescence, including fatty acids, such as stearic acid, and ceramide phosphoethanolamine species (CerPE). Similar to previous reports, alterations were detected in putative fatty acid building blocks, ceramides, among other lipid species. Altogether, our results reveal the ability to detect lipids implicated in senescence and show alterations to protein expression between normal and senescent fibroblast populations, including differences between young and aged cells. This report is the first time that the MALDI-IHC system has been utilized at a single-cell level to analyze both protein expression and lipid profiles in cultured cells, with a particular focus on changes associated with aging and senescence.

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来源期刊
CiteScore
5.50
自引率
9.40%
发文量
257
审稿时长
1 months
期刊介绍: The Journal of the American Society for Mass Spectrometry presents research papers covering all aspects of mass spectrometry, incorporating coverage of fields of scientific inquiry in which mass spectrometry can play a role. Comprehensive in scope, the journal publishes papers on both fundamentals and applications of mass spectrometry. Fundamental subjects include instrumentation principles, design, and demonstration, structures and chemical properties of gas-phase ions, studies of thermodynamic properties, ion spectroscopy, chemical kinetics, mechanisms of ionization, theories of ion fragmentation, cluster ions, and potential energy surfaces. In addition to full papers, the journal offers Communications, Application Notes, and Accounts and Perspectives
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