Methodology of determining the metabolomic profile of tumor-associated macrophages and monocytes in oncological diseases

Breast cancer is the leading cause of cancer-related death among women worldwide. Tumor-associated macrophages (TAMs) constitute primary component innate immunity in breast tissue. During development new approaches for treatment aimed at editing epigenome TAM, precise methods analysis macrophage metabolome are required to examine effect on metabolism. Our study develop an HPLC-MS/MS-based analytical approach characterize human immune cells (TAMs and their precursors, monocytes). Analysis lipid extracts was conducted a Dionex UltiMate 3000 liquid chromatograph connected Maxis Impact qTOF mass analyzer with ESI ion source. Quantitative 38 amino acids using Jasem Amino Acids LC-MS/MS Kit HPLC-MS/MS chromatographic system consisting out Agilent 6460 triple quadrupole spectrometric detector (Agilent), 1260 II (Agilent ) acids-HPLC Column (Jasem). The modified Folch method double extraction found be optimal approached sample preparation, since it enables simultaneously isolate extract water-soluble substances, particular, acids. reversed-phase chromatography yielded more useful data cell composition than hydrophilic interaction (HILIC). minimum number determine (TAM monocytes) identified as 2 × 106. Thus, we have developed acid modelled TAMs monocytes isolated patients minimal amount clinical material.