title: Secreted proteome of the murine multipotent hematopoietic progenitor cell line DKmix.
abstract: Administration of the multipotent hematopoietic progenitor cell (HPC) line DKmix improved cardiac function after myocardial infarction and accelerated dermal wound healing due to paracrine mechanisms. The aim of this study was to analyse the secreted proteins of DKmix cells in order to identify the responsible paracrine factors and assess their relevance to the wide spectrum of therapeutic effects. A mass spectrometry (MS)-based approach was used to identify secreted proteins of DKmix cells. Serum free culture supernatants of DKmix-conditioned medium were collected and the proteins present were separated, digested by trypsin and the resulting peptides were then analyzed by matrix-assisted laser desorption/ionization tandem time-of-flight (MALDI-TOF/TOF) MS. Overall 95 different proteins were identified. Among them, secretory proteins galectin-3 and gelsolin were identified. These proteins are known to stimulate cell migration and influence wound healing and cardiac remodelling. The remaining proteins originate from intracellular compartments like cytoplasm (69%), nucleus (12%), mitochondria (4%), and cytoplasmic membrane (3%) indicating permeable or leaky DKmix cells in the conditioned medium. Additionally, a sandwich immunoassay was used to detect and quantify cytokines and chemokines. Interleukin-6 (IL-6), interleukin-13 (IL-13), monocyte-chemoattractant protein-1 (MCP-1), monocyte-chemoattractant protein-3 (MCP-3), monocyte-chemoattractant protein-1alpha (MIP-1alpha) and monocyte-chemoattractant protein-1beta (MIP-1beta) were detected in low concentrations. This study identified a subset of proteins present in the DKmix-conditioned medium that act as paracrine modulators of tissue repair. Moreover, it suggests that DKmix-derived conditioned medium might have therapeutic potency by promoting tissue regeneration.
authors.affiliation: Hannover Medical School, Institute of Toxicology, Carl-Neuberg-Str. 1, 30625 Hannover, Germany.
authors.forename: Nina
authors.initials: N
authors.lastname: Luecke
authors.forename: Christian
authors.initials: C
authors.lastname: Templin
authors.forename: Marika Victoria
authors.initials: MV
authors.lastname: Muetzelburg
authors.forename: Detlef
authors.initials: D
authors.lastname: Neumann
authors.forename: Ingo
authors.initials: I
authors.lastname: Just
authors.forename: Andreas
authors.initials: A
authors.lastname: Pich
mesh_terms: D000818:Animals; D002460:Cell Line / Q000378:metabolism; D002465:Cell Movement / Q000187:drug effects; D017077:Culture Media, Conditioned; D016207:Cytokines / Q000737:chemistry; D005347:Fibroblasts / Q000378:metabolism; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D039902:Multipotent Stem Cells / Q000166:cytology / Q000378:metabolism*; D019899:Paracrine Communication; D010446:Peptide Fragments / Q000737:chemistry / Q000145:classification; D020543:Proteome / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D013347:Subcellular Fractions / Q000737:chemistry
chemical_list: D017077:Culture Media, Conditioned; D016207:Cytokines; D010446:Peptide Fragments; D020543:Proteome