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Updated: Jan 6, 2026

Isolation of Nuclei from Flash-Frozen Liver Tissue for Single-Cell Multiomics

Isolation of Nuclei from Flash-Frozen Liver Tissue for Single-Cell Multiomics

Published on: December 9, 2022

优化多组学样本准备:对 HepG2 细胞提取协议的比较评估.

Tilman F Arnst¹, Selina Hemmer¹, Claudia Fecher-Trost¹

¹Department of Experimental and Clinical Toxicology and Pharmacology, Institute of Experimental and Clinical Pharmacology and Toxicology, Center for Molecular Signaling (PZMS), PharmaScienceHub (PSH), Saarland University, Homburg, Germany.

Analytical and bioanalytical chemistry

|November 19, 2025

概括

这项研究比较了HepG2细胞的两种多组抽取方法. 使用偏磁珠的单相提取为同时进行代谢物,脂质和蛋白质分析提供了最可复制,最有效和最具成本效益的方法.

关键词:

在HepG2细胞中.质谱测量质量谱测量代谢学代谢学代谢学多种组合的多种组合.蛋白质组学是指蛋白质组学.样品的准备样品的准备

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Last Updated: Jan 6, 2026

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Published on: December 19, 2025

科学领域:

生物化学生物化学
蛋白质组学是指蛋白质组学.
代谢学代谢学代谢学
利皮多米克 (Lipidomics) 是一种消化剂.

背景情况:

多组学方法整合了来自多个分子层的数据,以进行全面的生物系统表征.
从单个样本同时生成多组数据对于最小化生物变异性和确保跨层一致性至关重要.
现有的多学科工作流程往往需要对特定的实验条件和仪器进行重大调整.

研究的目的:

系统地比较两个既定的协议,同时从HepG2细胞中提取代谢物,脂质和蛋白质.
为了评估珠子大小和消化条件在单相提取工作流程中的影响.
为了确定HepG2细胞系应用中最可复制,最有效和最具成本效益的多组分提取协议.

主要方法:

两相提取协议 (间相颗粒消化) 与单相提取协议 (珠子消化) 的比较.
通过调查珠子大小和消化条件来优化单相提取.
分析代谢学,脂质学和蛋白质学数据,使用液态染色学与高分辨率并列质谱 (LC-MS/MS) 和离子移动性分离相结合.

主要成果:

双相和单相协议均根据特征计数,选择性,可重复性,处理复杂性和整体性能进行了评估.
使用具有缩短潜伏时间的偏磁珠的单相提取证明了卓越的可重复性.
与双相方法相比,优化的单相方法被发现更有效和更具成本效益.

结论:

在所有评估标准中,没有一个提取协议是普遍最佳的.
使用具有优化消化条件的偏磁珠的单相提取方法为 HepG2 细胞的内部多态工作流提供了最有利的解决方案.
这种优化的协议提高了集成的OMIC数据生成的可重现性,效率和成本效益.