title: Proteomic analysis of tumor necrosis factor-alpha-induced secretome of human adipose tissue-derived mesenchymal stem cells.
abstract: Human adipose tissue-derived mesenchymal stem cells (hASCs) are useful for regeneration of inflamed or injured tissues. To identify secreted hASC proteins during inflammation, hASCs were exposed to tumor necrosis factor-alpha (TNF-alpha) and conditioned media derived from hASCs were analyzed by liquid chromatography coupled with tandem mass spectrometry. We identified 187 individual proteins as secreted proteins (secretome) in hASC-conditioned media; 118 proteins were secreted at higher levels upon TNF-alpha treatment. The TNF-alpha-induced secretome included a variety of cytokines and chemokines such as interleukin-6 (IL-6), IL-8, chemokine (C-X-C motif) ligand 6, and monocyte chemotactic protein-1 (MCP-1). TNF-alpha also increased expression of various proteases including cathepsin L, matrix metalloproteases and protease inhibitors, and induced secretion of long pentraxin 3, a key inflammatory mediator implicated in innate immunity. TNF-alpha-conditioned media stimulated migration of human monocytes, which play a key role in inflammatory responses. This migration was abrogated by pretreatment with neutralizing anti-IL-6, anti-IL-8, and anti-MCP-1 antibodies, suggesting that IL-6, IL-8, and MCP-1 are involved in migration of monocytes. Taken together, these results suggest that TNF-alpha-induced secretome may play a pivotal role in inflammatory responses and that shotgun proteomic analysis will be useful for elucidation of the paracrine functions of mesenchymal stem cells.
authors.affiliation: Medical Research Center for Ischemic Tissue Regeneration & Medical Research Institute, College of Medicine, Pusan National University, Yangsan, Republic of Korea.
authors.forename: Mi Jeong
authors.initials: MJ
authors.lastname: Lee
authors.forename: Jaeyoon
authors.initials: J
authors.lastname: Kim
authors.forename: Min Young
authors.initials: MY
authors.lastname: Kim
authors.forename: Yoe-Sik
authors.initials: YS
authors.lastname: Bae
authors.forename: Sung Ho
authors.initials: SH
authors.lastname: Ryu
authors.forename: Taehoon G
authors.initials: TG
authors.lastname: Lee
authors.forename: Jae Ho
authors.initials: JH
authors.lastname: Kim
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000704:Analysis of Variance; D002403:Cathepsins / Q000378:metabolism; D002465:Cell Movement / Q000502:physiology; D002853:Chromatography, Liquid; D016207:Cytokines / Q000378:metabolism; D015870:Gene Expression / Q000187:drug effects; D020869:Gene Expression Profiling; D006801:Humans; D007249:Inflammation / Q000378:metabolism; D020781:Matrix Metalloproteinase 1 / Q000378:metabolism; D059630:Mesenchymal Stem Cells / Q000187:drug effects / Q000378:metabolism*; D009000:Monocytes / Q000378:metabolism; D020543:Proteome / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D040901:Proteomics / Q000379:methods*; D015203:Reproducibility of Results; D053719:Tandem Mass Spectrometry; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism / Q000494:pharmacology*
chemical_list: D016207:Cytokines; D020543:Proteome; D014409:Tumor Necrosis Factor-alpha; D002403:Cathepsins; C522360:MMP1 protein, human; D020781:Matrix Metalloproteinase 1