Neonatal Research Group, Health Research Institute La Fe, Valencia, Spain.
Regenerative Medicine and Heart Transplantation Unit, Health Research Institute La Fe, Valencia, Spain.
Methods Mol Biol. 2023;2571:177-188. doi: 10.1007/978-1-0716-2699-3_18.
Extracellular vesicles (EVs) are secreted by cells and can be found in biological fluids (e.g., blood, saliva, urine, cerebrospinal fluid, and milk). EV isolation needs to be optimized carefully depending on the type of biofluid and tissue. Human milk (HM) is known to be a rich source of EVs, and they are thought to be partially responsible for the benefits associated with breastfeeding. Here, a workflow for the isolation and lipidomic analysis of HM-EVs is described. The procedure encompasses initial steps such as sample collection and storage, a detailed description for HM-EV isolation by multistage ultracentrifugation, metabolite extraction, and analysis by liquid chromatography coupled to mass spectrometry, as well as data analysis and curation.
细胞分泌的细胞外囊泡（EVs）存在于生物体液（如血液、唾液、尿液、脑脊液和乳汁）中。根据生物体液和组织的类型，需要仔细优化 EV 的分离。人乳（HM）是 EV 的丰富来源，并且被认为部分负责与母乳喂养相关的益处。这里描述了一种用于分离和脂质组学分析 HM-EV 的工作流程。该程序包括初始步骤，如样品采集和储存，通过多步超速离心详细描述 HM-EV 分离，代谢物提取，以及与质谱联用的液相色谱分析，以及数据分析和管理。
