title: Direct labeling of hMSC with SPIO: the long-term influence on toxicity, chondrogenic differentiation capacity, and intracellular distribution.
abstract: OBJECTIVE
  The purpose of this study was to evaluate the long-term cellular toxicity, labeling efficiency, chondrogenic differentiation capacity, and intracellular distribution following direct superparamagnetic iron oxide (SPIO) nanoparticle labeling of human mesenchymal stem cells (hMSCs) in the absence of transfection agents.
  
  
  METHODS
  hMSCs were incubated with a SPIO, Ferucarbotran, at concentrations of 0, 1, 10, and 100 μg Fe/ml for 24 or 72 h. The cell granularity and size change, reactive oxygen species generation, and mitochondria membrane potential were measured by flow cytometry. The differentiation capacity of the cells into chondrocytes was determined by Alcian blue and Safranin-O staining, immunocytochemical analysis, and reverse transcription polymerase chain reaction.
  
  
  RESULTS
  The intracellular distribution of the internalized particles was visualized via confocal microscopy. No significant difference was found in the toxicity of labeled cells relative to controls. Successful chondrogenesis of Ferucarbotran-labeled hMSCs was confirmed. The intracellular SPIO nanoparticles were located within the lysosomes.
  
  
  CONCLUSIONS
  In conclusion, we have demonstrated the feasibility of direct labeling with Ferucarbotran without impairment of cellular function, toxicity, or inhibition of differentiation capacity. Furthermore, lysosomal metabolism takes place after intracellular uptake of Ferucarbotran.
authors.affiliation: Institute of Biomedical Engineering, National Taiwan University, Taipei, Taiwan.|Department of Medical Imaging, National Taiwan University Hospital Yun-Lin Branch, Douliu, Taiwan.|Department of Medical Imaging, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan.
authors.forename: Chung-Yi
authors.initials: CY
authors.lastname: Yang
authors.affiliation: Department of Medical Imaging, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan.
authors.forename: Jong-Kai
authors.initials: JK
authors.lastname: Hsiao
authors.affiliation: Department of Physics, National Tsing Hua University, No. 101 Sec. 2, Kuang Fu Road, Hsinchu, 300, Taiwan.
authors.forename: Ming-Fong
authors.initials: MF
authors.lastname: Tai
authors.affiliation: Department of Biochemistry, Musculoskeletal Disease Center, J.L. Pettis VA Medical Center, Loma Linda University, Loma Linda, CA, 92357, USA.
authors.forename: Shin-Tai
authors.initials: ST
authors.lastname: Chen
authors.affiliation: Department of Medical Imaging, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan.
authors.forename: Hui-Ying
authors.initials: HY
authors.lastname: Cheng
authors.affiliation: Institute of Biomedical Engineering, National Taiwan University, Taipei, Taiwan.
authors.forename: Jaw-Lin
authors.initials: JL
authors.lastname: Wang
authors.affiliation: Department of Medical Imaging, National Taiwan University Hospital and National Taiwan University College of Medicine, Taipei, Taiwan. hmliu@ntu.edu.tw.|Department of Radiology, National Taiwan University Hospital and College of Medicine, 7 Chung-Shan South Road, Taipei, Taiwan. hmliu@ntu.edu.tw.
authors.forename: Hon-Man
authors.initials: HM
authors.lastname: Liu
mesh_terms: D002454:Cell Differentiation / Q000187:drug effects*; D020219:Chondrogenesis / Q000187:drug effects*; D003911:Dextrans / Q000633:toxicity*; D016326:Extracellular Matrix Proteins / Q000235:genetics / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D042541:Intracellular Space / Q000187:drug effects / Q000378:metabolism*; D058185:Magnetite Nanoparticles / Q000633:toxicity*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D018613:Microscopy, Confocal; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D013194:Staining and Labeling; D013997:Time Factors
chemical_list: D003911:Dextrans; D016326:Extracellular Matrix Proteins; D058185:Magnetite Nanoparticles; D012333:RNA, Messenger; D017382:Reactive Oxygen Species; C065507:ferumoxides