title: [Isolation and proteomic analysis of exosomes secreted by human cancer cells in vitro].
abstract: Exosomes are 20-100 nm membrane vesicles of endocytic origin secreted by most cell types in vitro and in vivo. Since exosomes contain both RNA (mRNA and microRNA) and proteins, which can be transferred to another cell, and be functional in that new environment, these vesicles may be involved in the communication between cells. The secretion of exosomes by tumor cells and their implication in the transport and propagation of infectious cargo suggest their participation in pathological situations. Our purpose here is to describe methods for the production, purification, and proteomic characterization of exosomes derived from human cancer cells in vitro. Based on exosomes' unique lipidic composition, we have developed the new approach to increase production of exosomes by cells in vitro. Secondly, we have developed quality control by laser correlation spectroscopy for exosomal assays based on the amount of MHC class I and CD63 molecules on their surface. At last, matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry was used after 2D electrophoresis for the proteomic analysis of exosomes derived from cancer cell lines. This study describes the protein composition of brain tumor cell-derived exosomes in more detail.
mesh_terms: D001692:Biological Transport; D015415:Biomarkers / Q000032:analysis; D045744:Cell Line, Tumor; D015180:Electrophoresis, Gel, Two-Dimensional; D055354:Exosomes / Q000737:chemistry* / Q000378:metabolism; D005910:Glioma / Q000737:chemistry* / Q000378:metabolism; D015395:Histocompatibility Antigens Class I / Q000032:analysis*; D006801:Humans; D009363:Neoplasm Proteins / Q000032:analysis*; D040901:Proteomics; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D060149:Tetraspanin 30 / Q000032:analysis*
chemical_list: D015415:Biomarkers; D015395:Histocompatibility Antigens Class I; D009363:Neoplasm Proteins; D060149:Tetraspanin 30