title: Mapping the secretome of human chondrogenic progenitor cells with mass spectrometry.
abstract: Tissue engineering offers promising perspectives in the therapy of osteoarthritis. In the context of cell-based therapy, chondrogenic progenitor cells (CPCs) may be used to regenerate defects in cartilage tissue. An in-depth characterization of the secretome of CPCs is a prerequisite to this approach. In this study, a method was developed for the qualitative and quantitative analysis of the secretome of undifferentiated and differentiated CPCs. Secreted proteins from cells grown in two-dimensional as well as three-dimensional alginate cultures were extracted and analyzed by liquid chromatography/tandem mass spectrometry (LC-MS/MS). Quantitation was achieved by internal standardization using stable isotope-labeled amino acids in cell culture (SILAC). Qualitative analysis of CPC secretomes revealed ECM-components, signal proteins and growth factors most of which were also found in healthy cartilage. A quantitative comparison revealed significantly upregulated proteins with regenerative potential during differentiation, while proteins involved in catabolic metabolism were significantly downregulated. The development of methods for qualitative and quantitative analysis of the secretome of CPCs by mass spectrometry provides a foundation for the investigation of progenitor or stem cells from other sources.
authors.affiliation: University Medical Center Göttingen, Department of Orthodontics, Robert-Koch-Straße 40, 37075 Göttingen, Germany; University Medical Center Göttingen, Tissue Regeneration Work Group, Robert-Koch-Straße 40, 37075 Göttingen, Germany. Electronic address: sarah.batschkus@med.uni-goettingen.de.
authors.forename: Sarah
authors.initials: S
authors.lastname: Batschkus
authors.affiliation: Max Planck Institute for Biophysical Chemistry, Biophysical Mass Spectrometry Group, Am Fassberg 11, 37077 Göttingen, Germany.
authors.forename: Ilian
authors.initials: I
authors.lastname: Atanassov
authors.affiliation: Max Planck Institute for Biophysical Chemistry, Biophysical Mass Spectrometry Group, Am Fassberg 11, 37077 Göttingen, Germany; University Medical Center Göttingen, Institute of Clinical Chemistry, Bioanalytics Group, Robert-Koch-Straße 40, 37075 Göttingen, Germany.
authors.forename: Christof
authors.initials: C
authors.lastname: Lenz
authors.affiliation: University Medical Center Göttingen, Department of Orthodontics, Robert-Koch-Straße 40, 37075 Göttingen, Germany.
authors.forename: Philipp
authors.initials: P
authors.lastname: Meyer-Marcotty
authors.affiliation: University Medical Center Göttingen, Department of Orthodontics, Robert-Koch-Straße 40, 37075 Göttingen, Germany; University Medical Center Göttingen, Tissue Regeneration Work Group, Robert-Koch-Straße 40, 37075 Göttingen, Germany.
authors.forename: Gökhan
authors.initials: G
authors.lastname: Cingöz
authors.affiliation: University Medical Center Regensburg, Department of Orthodontics, Franz-Josef-Strauß-Allee 11, 93053 Regensburg, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Kirschneck
authors.affiliation: Max Planck Institute for Biophysical Chemistry, Biophysical Mass Spectrometry Group, Am Fassberg 11, 37077 Göttingen, Germany; University Medical Center Göttingen, Institute of Clinical Chemistry, Bioanalytics Group, Robert-Koch-Straße 40, 37075 Göttingen, Germany.
authors.forename: Henning
authors.initials: H
authors.lastname: Urlaub
authors.affiliation: University Medical Center Göttingen, Tissue Regeneration Work Group, Robert-Koch-Straße 40, 37075 Göttingen, Germany.
authors.forename: Nicolai
authors.initials: N
authors.lastname: Miosge
mesh_terms: D000328:Adult; D000818:Animals; D002358:Cartilage, Articular / Q000033:anatomy & histology / Q000166:cytology / Q000473:pathology*; D002417:Cattle; D002478:Cells, Cultured; D011232:Chemical Precipitation; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005110:Extracellular Space / Q000737:chemistry; D051445:Fibrocartilage / Q000166:cytology; D006801:Humans; D010003:Osteoarthritis / Q000473:pathology* / Q000628:therapy; D011506:Proteins / Q000032:analysis / Q000302:isolation & purification / Q000378:metabolism*; D020543:Proteome / Q000737:chemistry* / Q000378:metabolism; D013234:Stem Cells / Q000737:chemistry / Q000378:metabolism*; D053719:Tandem Mass Spectrometry
chemical_list: D011506:Proteins; D020543:Proteome
keywords: Chondrogenic progenitor cells; Mass spectrometry; Osteoarthritis; Secretome; Tissue engineering