Metabolomic profiling of rare cell populations isolated by flow cytometry from tissues

• DOI: 10.7554/elife.61980
• Publication Date: 2021-01-20T13:02:48Z
• Authors: Andrew W DeVilbiss; Zhiyu Zhao; Misty S Martin-Sa...; Jessalyn M Ubella...; Alpaslan Tasdogan; Michalis Agathocl...; Thomas P Mathews; Sean J Morrison
• Keywords: Metabolome; Purine metabolism; Metabolic pathway

Little is known about the metabolic regulation of rare cell populations because most metabolites are hard to detect in small numbers cells. We previously described a method for metabolomic profiling flow cytometrically isolated hematopoietic stem cells (HSCs) that detects 60 10,000 (Agathocleous et al., 2017). Here we describe new involving hydrophilic liquid interaction chromatography and high-sensitivity orbitrap mass spectrometry detected 160 HSCs, including many more glycolytic lipid intermediates. improved chromatographic separation, increased resolution, minimized ion suppression, eliminated sample drying. Most metabolite levels did not significantly change during isolation. Mouse HSCs exhibited glycerophospholipids relative bone marrow methotrexate treatment altered purine biosynthesis. Circulating human melanoma were depleted intermediates subcutaneous tumors, suggesting decreased synthesis metastasis. These methods facilitate routine analysis from tissues.