Identification and Characterization of 293T Cell-Derived Exosomes by Profiling the Protein, mRNA and MicroRNA Components

PLoS One. 2016 Sep 20;11(9):e0163043. doi: 10.1371/journal.pone.0163043. eCollection 2016.

Abstract

Cell-derived exosomes are leading candidates for in vivo drug delivery carriers. In particular, exosomes derived from 293T cells are used most frequently, although exosome dosing has varied greatly among studies. Considering their biological origin, it is crucial to characterize the molecular composition of exosomes if large doses are to be administered in clinical settings. In this study, we present the first comprehensive analysis of the protein, messenger RNA and microRNA profiles of 293T cell-derived exosomes; then, we characterized these data using Gene Ontology annotation and Kyoto Encyclopedia for Genes and Genomes pathway analysis. Our study will provide the basis for the selection of 293T cell-derived exosome drug delivery systems. Profiling the exosomal signatures of 293T cells will lead to a better understanding of 293T exosome biology and will aid in the identification of any harmful factors in exosomes that could cause adverse clinical effects.

MeSH terms

  • Blotting, Western
  • Chromatography, Liquid
  • Exosomes / genetics*
  • Exosomes / metabolism*
  • Exosomes / ultrastructure
  • Gene Expression Profiling / methods*
  • Gene Ontology
  • HEK293 Cells
  • Humans
  • MicroRNAs / genetics
  • Microscopy, Electron, Transmission
  • Oligonucleotide Array Sequence Analysis / methods
  • Proteins / metabolism
  • Proteomics / methods*
  • RNA, Messenger / genetics
  • Tandem Mass Spectrometry

Substances

  • MicroRNAs
  • Proteins
  • RNA, Messenger

Grants and funding

National Basic Research Program of China (973 Program) (No. 2011CB504803), Natural Science Foundation of Jiangsu Province, China (BK20130890, BK2011013 and BK2012014), the National Natural Science Foundation of China (31200969, 31200874, 81101330, 31271378 and 81250044), the National Science Fund for Distinguished Young Scholars (81025019) and the program for New Century Excellent Talents in University from the Ministry of Education, China (NCET-12-0261). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.