title: Evaluation of metabolomic changes as a biomarker of chondrogenic differentiation in 3D-cultured human mesenchymal stem cells using proton (1H) nuclear magnetic resonance spectroscopy.
abstract: OBJECTIVE
  The purpose of this study was to evaluate the metabolomic changes in 3D-cultured human mesenchymal stem cells (hMSCs) in alginate beads, so as to identify biomarkers during chondrogenesis using (1)H nuclear magnetic resonance (NMR) spectroscopy.
  
  
  METHODS
  hMSCs (2×10(6) cells/mL) were seeded into alginate beads, and chondrogenesis was allowed to progress for 15 days. NMR spectra of the chondrogenic hMSCs were obtained at 4, 7, 11, and 15 days using a 14.1-T (600-MHz) NMR with the water suppression sequence, zgpr. Real-Time polymerase chain reaction (PCR) was performed to confirm that that the hMSCs differentiated into chondrocytes and to analyze the metabolomic changes indicated by the NMR spectra.
  
  
  RESULTS
  During chondrogenesis, changes were detected in several metabolomes as hMSC chondrogenesis biomarkers, e.g., fatty acids, alanine, glutamate, and phosphocholine. The metabolomic changes were compared with the Real-Time PCR results, and significant differences were determined using statistical analysis. We found that changes in metabolomes were closely related to biological reactions that occurred during the chondrogenesis of hMSCs.
  
  
  CONCLUSIONS
  In this study, we confirm that metabolomic changes detected by (1)H-NMR spectroscopy during chondrogenic differentiation of 3D-cultured hMSCs in alginate beads can be considered as biomarkers of stem cell differentiation.
authors.affiliation: Department of Biomedical Engineering/UHRC, Inje University, Gimhae, Gyeongnam, South Korea.
authors.forename: Moo-Young
authors.initials: MY
authors.lastname: Jang
authors.forename: Song-I
authors.initials: SI
authors.lastname: Chun
authors.forename: Chi-Woong
authors.initials: CW
authors.lastname: Mun
authors.forename: Kwan Soo
authors.initials: KS
authors.lastname: Hong
authors.forename: Jung-Woog
authors.initials: JW
authors.lastname: Shin
mesh_terms: D015415:Biomarkers / Q000378:metabolism*; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D019902:Chondrocytes / Q000378:metabolism / Q000502:physiology; D020219:Chondrogenesis / Q000502:physiology*; D006801:Humans; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D055442:Metabolome / Q000502:physiology*; D055432:Metabolomics / Q000379:methods; D011522:Protons
chemical_list: D015415:Biomarkers; D011522:Protons